Formulation Library: 101 Buffer and Reagent Recipes

Citrate Buffer (pH 3.0 to 6.2)

Adjustable formulation, pH 3–6.2 · amounts depend on target pH — use the interactive calculator

Citrate buffers can be used for RNA isolation, due to its ability to prevent base hydrolysis. The buffer is also used for antigen detection by breaking cross-links between antigens and any substances in its fixation medium. Citrate buffer is a popular choice for immunofluorescent staining applications, as it can aid in stain intensity without contributing to background signal. It is a popular choice for biological research and clinical applications, including pathology, and has had an increasing role in the medical field, as it has been shown to mildly improve dialysis efficiency. It has a shelf life of up to 3 months at room temperature.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add Sodium Citrate dihydrate of Sodium Citrate dihydrate to the solution
3. Add Citric Acid of Citric Acid to the solution
4. Adjust solution to final desired pH using HCl or NaOH
5. Add distilled water until the volume is 1 L.

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Acetate Buffer (pH 3.6 to 5.6)

Adjustable formulation, pH 3.6–5.6 · amounts depend on target pH — use the interactive calculator

Sodium acetate buffers are used for purification and precipitation of nucleic acids, as well as for protein crystallization and staining gels used in protein electrophoresis. It is very popular in hematology, since there is some evidence that acetate-buffered infusions show improved stability. Acetate buffers are inexpensive and simple to prepare, and can be stored at room temperature.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add Sodium Acetate of Sodium Acetate to the solution
3. Add Acetic Acid of Acetic Acid to the solution
4. Adjust solution to final desired pH using HCl or NaOH
5. Add distilled water until the volume is 1 L.

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Tris Buffer (1 M, pH 7.2)

Fixed formulation, amounts given per 1 L of final solution

Tricine is derived from the amino acids tris and glycine. It is a dipolar ion (Zwitterionic) and hydroxyl radical scavenger, and is used extensively for SDS-PAGE applications for small proteins. Tricine buffer is also commonly used for electrophoresis procedures.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 121.14 g of Tris base to the solution
3. Adjust solution to desired pH using HCl (typically pH ≈ 7.0).
4. Add distilled water until the volume is 1 L.

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Citrate Buffer (0.1 M, pH 6.0)

Fixed formulation, amounts given per 1 L of final solution

Citrate buffers can be used for RNA isolation, thanks to its ability to prevent base hydrolysis. It also finds use in antigen detection by breaking cross-links between antigens and any substances in its fixation medium.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 24.269 g of Sodium Citrate dihydrate to the solution
3. Add 3.358 g of Citric Acid to the solution
4. Adjust solution to desired pH using 0.1N HCl (typically pH ≈ 6.0).
5. Add distilled water until the volume is 1 L.

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Acetate Buffer (0.1 M, pH 5.0)

Fixed formulation, amounts given per 1 L of final solution

Sodium acetate buffers are used for purification and precipitation of nucleic acids, as well as for protein crystallization and staining gels used in protein electrophoresis.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 5.772 g of Sodium Acetate to the solution
3. Add 1.778 g of Acetic Acid to the solution
4. Adjust solution to desired pH using 10N HCl (typically pH ≈ 5.0).
5. Add distilled water until the volume is 1 L.

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Phosphate Buffer (pH 5.8 to 7.4)

Adjustable formulation, pH 5.8–7.4 · amounts depend on target pH — use the interactive calculator

A simple phosphate buffer is used ubiquitously in biological experiments, as it can be adapted to a variety of pH levels, including isotonic. This wide range is due to phosphoric acid having 3 dissociation constants, (known in chemistry as a triprotic acid) allowing for formulation of buffers near each of the pH levels of 2.15, 7.21, or 12.32. Phosphate buffer is highly water soluble and has a high buffering capacity, but will inhibit enzymatic activity and precipitates in ethanol. The buffer is one of the most popular currently used, and is commonly employed in molecular and cell biology, chemistry, and material science, among many others.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add Sodium Phosphate Dibasic Heptahydrate of Sodium Phosphate Dibasic Heptahydrate to the solution
3. Add Sodium Phosphate Monobasic Monohydrate of Sodium Phosphate Monobasic Monohydrate to the solution
4. Adjust solution to final desired pH using HCl or NaOH
5. Add distilled water until the volume is 1 L.

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Hydrochloric Acid-Potassium Chloride Buffer (0.1 M, pH 2.0)

Fixed formulation, amounts given per 1 L of final solution

This common buffer is used in multiple biology research fields, often for enzymatic or histochemical usage. It should ideally be prepared shortly before use, as it does not store well.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 7.45 g of Potassium Chloride to the solution
3. Add 0.772 g of Hydrochloric acid to the solution
4. Adjust solution to final desired pH using HCl or NaOH
5. Add distilled water until the volume is 1 L.

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Glycine-HCl Buffer (0.1 M, pH 3.0)

Fixed formulation, amounts given per 1 L of final solution

This low-pH buffer is used extensively with affinity chromatography, particularly with proteins and antibodies. It is also used for cell culture and other biological applications.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 7.5 g of Glycine to the solution
3. Add 0.832 g of Hydrochloric acid to the solution
4. Adjust solution to final desired pH using HCl or NaOH
5. Add distilled water until the volume is 1 L.

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Citrate-Phosphate Buffer (0.15 M, pH 5.0)

Fixed formulation, amounts given per 1 L of final solution

A traditional buffer originally introduced in 1921. Since the Citrate-Phosphate buffer (also known as the McIlvaine buffer) only has 2 ingredients, the recipe can be adjusted to a pH range of 3-8. It is used for multiple applications in cell biology, molecular biology, and hematology.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 18.15 g of Sodium Phosphate Dibasic Dihydrate to the solution
3. Add 9.605 g of Citric Acid to the solution
4. Adjust solution to final desired pH using HCl or NaOH
5. Add distilled water until the volume is 1 L.

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Glycine-Sodium Hydroxide Buffer (0.08 M, pH 10)

Fixed formulation, amounts given per 1 L of final solution

A common buffer with good buffering capacity, often used in cellular biology. This buffer will slightly inhibit alkaline phosphatase enzymatic activity due to the configuration of the glycine amino group at higher pH levels. Due to this inhibition, it is recommended to use horseradish peroxidase (HRP) conjugates with this buffer.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 6.01 g of Glycine to the solution
3. Add 2.05 g of Sodium Hydroxide to the solution
4. Adjust solution to final desired pH using HCl or NaOH
5. Add distilled water until the volume is 1 L.

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EBSS (magnesium, calcium, phenol red) (pH 7.0)

Fixed formulation, amounts given per 1 L of final solution

EBSS is the abbreviation of Earle's Balanced Salt Solution, and is in common use for cell culture applications including washing and dilution in CO2 environments. The addition of the magnesium and calcium ions, along with phenol red, is to assist in reagent preparation. Phenol red is a weak estrogen and should not be used long-term with responsive cell cultures. For other buffers containing Phenol Red, click here.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 0.2 g of Calcium Chloride to the solution
3. Add 0.2 g of Magesium Sulfate Heptahydrate to the solution
4. Add 0.4 g of Potassium Chloride to the solution
5. Add 2.2 g of Sodium bicarbonate to the solution
6. Add 6.8 g of Sodium chloride to the solution
7. Add 0.14 g of Sodium Phosphate Monobasic Monohydrate to the solution
8. Add 1 g of D-Glucose (Dextrose) to the solution
9. Add 0.01 g of Phenol Red to the solution
10. Adjust solution to final desired pH using HCl or NaOH
11. Add distilled water until the volume is 1 L.

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Imidazole-HCl Buffer (0.05 M, pH 7.0)

Fixed formulation, amounts given per 1 L of final solution

Imidazole is a mild buffer that is useful for IHC applications in multiple research areas such as biochemistry and molecular biology. It has a good shelf-life and is well-tolerated by multiple cell types.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 3.404 g of Imidazole to the solution
3. Add 0.886 g of Hydrochloric acid to the solution
4. Adjust solution to desired pH using HCl (typically pH ≈ 7.0).
5. Add distilled water until the volume is 1 L.

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Trizma® Buffer (pH 7.0 to 9.2)

Adjustable formulation, pH 7–9.2 · amounts depend on target pH — use the interactive calculator

Trizma® is a proprietary chemical buffer used similarly to Tris buffer. It is commonly used in protein extraction for many types of IHC assays as well as blot applications. It is used in sandwich ELISA protocols for protection of the antibodies, and in electrophoresis of nucleic acids.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add Trizma Hydrochloride of Trizma Hydrochloride to the solution
3. Add Trizma Base of Trizma Base to the solution
4. Add distilled water until the volume is 1 L.

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MES (0.5 M, pH 6)

Fixed formulation, amounts given per 1 L of final solution

MES buffer is a Good's buffer that is remarkably stable both chemically and enzymatically. It has minimal UV absorbance and is often used as a running buffer for bis-tris gels.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 97.62 g of MES free acid to the solution
3. Starting pH for 0.5M MES solution is 3.23 For 1 L of 0.5 M MES, 13.6 ml of 10N NaOH is needed to adjust the pH to 6.0
4. Add dH2O until the volume is 1 L.

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Bis-Tris Buffer (1 M, 6.5 pH)

Fixed formulation, amounts given per 1 L of final solution

Bis-Tris buffer is used in molecular biology for applications involving both proteins and nucleic acids. Similar to tris-buffers, it has good buffer capacity and can be stored at room temperature. Bis-Tris interacts with several metal ions including Cu and Pb.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 209.24 g of Bis-Tris to the solution
3. Starting pH without HCl is 9.90. Concentrated HCl is used to adjust the pH. Filter sterilize or autoclave before use or storage.
4. Add dH2O until the volume is 1 L.

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ADA Buffer (0.5 M, 6.6 pH)

Fixed formulation, amounts given per 1 L of final solution

ADA buffer is one of the Good's buffer with a pH range of 6.0-7.2. It is often used in cell culture media as a chelating agent for H+, Ca2+, and Mg2+.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 95.11 g of ADA to the solution
3. Add dH2O until the volume is 1 L.
4. ADA will not fully dissolve until the pH is raised. Adjust solution to desired pH by 10N NaOH.

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ACES Buffer (0.1 M, 6.7 pH)

Fixed formulation, amounts given per 1 L of final solution

ACES buffer is one of the Good's zwitterionic buffer with a pH range of 6.1-7.2.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 18.22 g of ACES to the solution
3. Adjust solution to desired pH by 10N NaOH.
4. Add dH2O until the volume is 1 L.

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PIPES Buffer (1 M, 6.8 pH)

Fixed formulation, amounts given per 1 L of final solution

PIPES buffer is one of the Good's zwitterionic buffer with a pH range of 6.1-7.5.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 302.37 g of PIPES to the solution
3. Solubility of PIPES free acid is low in acidic range. Adjust solution to desired pH by 10N NaOH.
4. Add dH2O until the volume is 1 L.

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MOPSO Buffer (0.5 M, 6.9 pH)

Fixed formulation, amounts given per 1 L of final solution

MOPSO buffer is one of the Good's zwitterionic buffer with an approximate pH range of 6.5-7.9. It has a good buffering capacity but does react with iron ions. MOPSO is used for a variety of assays that require physiological pH.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 112.63 g of MOPSO, Free acid to the solution
3. Adjust solution to desired pH by 10N NaOH.
4. Add dH2O until the volume is 1 L.

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BES-Buffered Saline (2X) (0.05 M, pH 6.95)

Fixed formulation, amounts given per 1 L of final solution

BES-buffered saline is often used in the calcium phosphate-mediated transformation of mammalian cells.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 10.66 g of BES to the solution
3. Add 16.36 g of Sodium chloride to the solution
4. Add 0.21 g of Sodium Phosphate Dibasic to the solution
5. Adjust solution to desired pH using 1M NaOH (typical pH = 6.95)
6. Add dH2O until the volume is 1 L.

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TES Buffer (1 M, 7.5 pH)

Fixed formulation, amounts given per 1 L of final solution

TES buffer is one of the Good's zwitterionic buffer with a pH range of 6.8-8.2.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 229.25 g of TES to the solution
3. Adjust solution to desired pH by 10N NaOH.
4. Add dH2O until the volume is 1 L.

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MOPS Buffer (10X) (0.2 M, pH 7)

Fixed formulation, amounts given per 1 L of final solution

MOPS buffer is often used in polyacrylamide gel electrophoresis.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 41.86 g of MOPS free acid to the solution
3. Add 4.1 g of Sodium Acetate to the solution
4. Add 3.72 g of Disodium EDTA to the solution
5. Adjust solution to desired pH using NaOH (typical pH = 7)
6. Add dH2O until the volume is 1 L.

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SSC buffer (20X) (3 M, pH 7)

Fixed formulation, amounts given per 1 L of final solution

Saline-sodium citrate (SSC) buffer is often used as a hybridization buffer for DNA and RNA, particularly for Northern and Southern blots.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 175.3 g of Sodium chloride to the solution
3. Add 88.3 g of Sodium citrate to the solution
4. Adjust solution to desired pH with 14 N HCl (typically 7.0).
5. Add dH2O until the volume is 1 L.

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TBS (1 M, pH 7.4)

Fixed formulation, amounts given per 1 L of final solution

Tris-Buffered Saline (TBS) is a popular isotonic buffer used for multiple applications, including as a washing buffer in immunoassays of all kinds. It is one of the default buffers for antibody preparation. For other antibody preparation materials, click here.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 8 g of Sodium chloride to the solution
3. Add 0.2 g of Potassium Chloride to the solution
4. Add 3 g of Tris base to the solution
5. Adjust solution to desired pH with HCl (typically 7.4).
6. Add dH2O until the volume is 1 L.

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Bicine (1 M, pH 8.26)

Fixed formulation, amounts given per 1 L of final solution

Bicine is used in peptide and protein biochemical applications, and is ideal for low temperatures. Bicine is a Zwitterionic compound, also known as a dipolar ion.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 163.17 g of Bicine to the solution
3. Adjust solution to desired pH with 10N NaOH.
4. Add dH2O until the volume is 1 L.

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Tricine (1 M, pH 8.05)

Fixed formulation, amounts given per 1 L of final solution

Tricine is derived from the amino acids tris and glycine. It is a dipolar ion (Zwitterionic) and hydroxyl radical scavenger, and is used extensively for SDS-PAGE applications for small proteins. Tricine buffer is also commonly used for electrophoresis procedures.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 179.17 g of Tricine to the solution
3. Adjust solution to desired pH with 10N NaOH.
4. Add dH2O until the volume is 1 L.

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TAE (1 M, pH 8.6)

Fixed formulation, amounts given per 1 L of final solution

TAE buffer is commonly used with nucleic acids for agarose electrophoresis applications. For the associated fluorescent probes for gel electrophoresis, click here.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 242 g of Tris base to the solution
3. Add 18.61 g of Disodium EDTA to the solution
4. Add 59.955 g of Acetic Acid to the solution
5. The 1x TAE solution is 40mM Tris, 20mM Acetate and 1mM EDTA and typically has a pH around 8.6 (do not adjust).
6. Add dH2O until the volume is 1 L.

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CAPS (0.5 M, pH 10.4)

Fixed formulation, amounts given per 1 L of final solution

CAPS is used commonly for Western Blot applications and other protein sorting procedures. The very high pH of this buffer allows for identification and sequencing of peptides and proteins with high isoelectric points (pl or IEP).

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 110.66 g of CAPS to the solution
3. Adjust solution to desired pH using 10N NaOH.
4. Add dH2O until the volume is 1 L.

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HEPPSO (1 M, pH 7.85)

Fixed formulation, amounts given per 1 L of final solution

HEPPSO is a Good's buffer used with protein separation in biological research. It is a Zwitterionic compound and binds strongly with copper ions.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 268.33 g of HEPPSO to the solution
3. Adjust solution to desired pH with 10N NaOH.
4. Add dH2O until the volume is 1 L.

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CHES (0.5 M, pH 9.5)

Fixed formulation, amounts given per 1 L of final solution

CHES is a dipolar ion (Zwitterionic) used at relatively high pH levels up to 10. It is employed in several biological research fields including enzymology, where it is useful for studying reactions above physiological pH range. Commonly used for electrophoresis, it has low reactivity with most metal ions.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 103.65 g of CHES to the solution
3. Adjust solution to desired pH with 10N NaOH.
4. Add dH2O until the volume is 1 L.

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Potassium Phosphate (pH 5.8 to 8.0)

Adjustable formulation, pH 5.8–8 · amounts depend on target pH — use the interactive calculator

Potassium phosphate buffers, sometimes called Gomori buffers, consist of a mixture of monobasic dihydrogen phosphate and dibasic monohydrogen phosphate. These buffers have excellent buffering capacity and are highly soluble in water. However, potassium phosphate buffer reacts with some metal ions including calcium and magnesium, and inhibits enzymatic reactions. Additionally, all phosphates precipitate in ethanol, which can be deleterious to some nucleic acid preparations that typically use ethanol solutions. Potassium phosphate buffer is used in research fields ranging from molecular biology to botany, due to its adaptability and affordability.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add Potassium phosphate dibasic of Potassium phosphate dibasic to the solution
3. Add Potassium Phosphate Monobasic of Potassium Phosphate Monobasic to the solution
4. Add dH2O until the volume is 1 L.

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Diethanolamine (1 M, pH 9.8)

Fixed formulation, amounts given per 1 L of final solution

Diethanolamine (DEA) is the buffer of choice in applications employing concentrated Tropix substrates for the detection of alkaline phosphatase. DEA buffers provide favorable reaction kinetics and greater sensitivity compared to most buffer systems.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 105.14 g of Diethanolamine to the solution
3. Add 0.1 g of Magnesium Chloride Hexahydrate to the solution
4. Adjust the pH to 9.8 with 10 M HCl solution.
5. Add dH2O until the volume is 1 L.

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Sodium Borate Buffer (1 M, pH 8.5)

Fixed formulation, amounts given per 1 L of final solution

Sodium Borate Buffer is a buffer that is used to maintain the pH within a relatively narrow range. It is isotonic and has a strong bactericidal effect, and can be used to dilute substances or in coating procedures.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 61.83 g of Boric Acid (H3BO3) to the solution
3. Add 10 g of Sodium Hydroxide to the solution
4. Add dH2O until the volume is 1 L.

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Maleic Acid (1 M, pH 7.5)

Fixed formulation, amounts given per 1 L of final solution

Maleic acid buffer is commonly used for Southern blot preparation and as a washing buffer in numerous applications involving nucleic acids. It has a shelf life of 6 months to a year.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 116.1 g of Maleic Acid to the solution
3. Add 87.6 g of Sodium chloride to the solution
4. Add 40 g of Sodium Hydroxide to the solution
5. Add dH2O until the volume is 1 L.
6. Sterilize by autoclaving at 15 psi for 20 minutes or by filtration. Store at room temperature. For washing applications, add 0.2% Tween-20 if desired.

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Tris-Buffered Saline (TBS, 0.1 M)

Fixed formulation, amounts given per 1 L of final solution

Tris-Buffered Saline (TBS) is commonly used for washing in many applications, such as cell culture, IHC, WB, and ELISA.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 8 g of Sodium chloride to the solution
3. Add 0.2 g of Potassium Chloride to the solution
4. Add 12.1 g of Tris to the solution
5. Adjust the pH to 7.4 with HCl.
6. Add distilled water until the volume is 1 L.

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Ammonium Sulfate, Saturated

Fixed formulation, amounts given per 1 L of final solution

Ammonium Sulfate is commonly used for protein precipitation and fractionation due to enhancing hydrophobic interactions.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 761 g of ammonium sulfate to the solution
3. Add distilled water until the volume is 1 L.
4. Dissolve with stirring and gentle heating. Cool to room temperature and adjust the pH to 7 (with NH4OH or an acid such as HCl). If excess is to be stored, sterilize the solution by filtering through a 0.2-µm filter and store at 4°C.

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Potassium Phosphate Buffer (1 M, pH 6.5)

Fixed formulation, amounts given per 1 L of final solution

Potassium phosphate buffers, sometimes called Gomori buffers, consist of a mixture of monobasic dihydrogen phosphate and dibasic monohydrogen phosphate. These buffers have excellent buffering capacity and are highly soluble in water. However, potassium phosphate buffer reacts with some metal ions including calcium and magnesium, and inhibits enzymatic reactions. Additionally, all phosphates precipitate in ethanol, which can be deleterious to some nucleic acid preparations that typically use ethanol solutions. Potassium phosphate buffer is used in research fields ranging from molecular biology to botany, due to its adaptability and affordability.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 95 g of Potassium Phosphate Monobasic to the solution
3. Add 52.5 g of Potassium phosphate dibasic to the solution
4. Adjust the pH to 6.5.
5. Add distilled water until the volume is 1 L.
6. Filter sterilize and store at room temperature.

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Citrate-Phosphate Buffer (110 mM, pH 5.6)

Fixed formulation, amounts given per 1 L of final solution

A traditional buffer originally introduced in 1921. Since the Citrate-Phosphate buffer (also known as the McIlvaine buffer) only has 2 ingredients, the recipe can be adjusted to a pH range of 3-8. It is used for multiple applications in cell biology, molecular biology, and hematology.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 7.1 g of Sodium phosphate dibasic (anhydrous) to the solution
3. Add 11.5 g of Citric Acid to the solution
4. Add distilled water until the volume is 1 L.
5. Sterilize by autoclaving. (The final solution contains 50 mM Na2HPO4.)

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Citrate-Phosphate Buffer (40 mM, pH 5.6)

Fixed formulation, amounts given per 1 L of final solution

A traditional buffer originally introduced in 1921. Since the Citrate-Phosphate buffer (also known as the McIlvaine buffer) only has 2 ingredients, the recipe can be adjusted to a pH range of 3-8. It is used for multiple applications in cell biology, molecular biology, and hematology.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 2.82 g of Sodium phosphate dibasic (anhydrous) to the solution
3. Add 4.2 g of Citric Acid to the solution
4. Add distilled water until the volume is 1 L.
5. Sterilize by autoclaving. (The final solution contains 20 mM Na2HPO4.)

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Sodium Phosphate Solution

Fixed formulation, amounts given per 1 L of final solution

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 70 g of Sodium Phosphate Dibasic to the solution
3. Add 30 g of Sodium Phosphate Monobasic Monohydrate to the solution
4. Add distilled water until the volume is 1 L.
5. Autoclave and store at room temperature.

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Sodium Acetate (3 M, pH 5.2)

Fixed formulation, amounts given per 1 L of final solution

Sodium Acetate (3 M) is commonly used during DNA and RNA extraction. Sodium acetate, which is a salt, helps precipitate nucleic acids.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 246.1 g of Sodium Acetate to the solution
3. Adjust the pH to 5.2 with glacial acetic acid. Allow the solution to cool overnight. Adjust the pH once more to 5.2 with glacial acetic acid.
4. Add distilled water until the volume is 1 L.
5. Filter-sterilize the solution.

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TB Buffer

Fixed formulation, amounts given per 1 L of final solution

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 0.37 g of EDTA to the solution
3. Add 13.64 g of Potassium Chloride to the solution
4. Add 2.74 g of Sodium chloride to the solution
5. Add 0.17 g of Phenylmethylsulfonyl fluoride (PMSF) to the solution
6. Add 1.21 g of Tris-hydrochloride to the solution
7. Adjust pH to 6.8.
8. Add distilled water until the volume is 1 L.
9. Sterilize by filtration.

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Tris-citrate Buffer (pH 8.0)

Fixed formulation, amounts given per 1 L of final solution

Tris-citrate buffer can be used for electrophoresis of bcterial enzymes.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 16 g of Citric Acid to the solution
3. Add 40.2 g of Tris to the solution
4. Add distilled water until the volume is 1 L.

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Borate Buffer

Fixed formulation, amounts given per 1 L of final solution

Borate buffer is a widely use buffer with many applications. Borate buffers that contain sodium chloride have been used to study corrosion.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 6.185 g of Boric acid to the solution
3. Add 4.385 g of Sodium chloride to the solution
4. Add 5.0305 g of Sodium tetraborate to the solution
5. Add distilled water until the volume is 1 L.
6. Use a stir bar and low heat to ensure that the boric acid dissolves fully. Sterilize by vacuum-filtration with a 0.22-µm filter. Adjust the pH to 8.4.

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Glycine (0.1 M, pH 2.2)

Fixed formulation, amounts given per 1 L of final solution

Glycine is a bulking agent that can help promote protein stability.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 7.5 g of Glycine (Sigma G7126) to the solution
3. Adjust the pH to 2.2 with HCl.
4. Add distilled water until the volume is 1 L.
5. Autoclave.

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Ammonium Bicarbonate (50 mM, pH 7.8)

Fixed formulation, amounts given per 1 L of final solution

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 4 g of Ammonium bicarbonate to the solution
3. Add distilled water until the volume is 1 L.

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Ammonium Acetate

Fixed formulation, amounts given per 1 L of final solution

Ammonium acetate is widely used in molecular biology in applications such as protein purification and precipitation.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 770 g of ammonium acetate to the solution
3. Add distilled water until the volume is 1 L.
4. Sterilize the solution by passing it through a 0.22-µm filter. Store the solution in tightly sealed bottles at 4°C or at room temperature. Ammonium acetate decomposes in hot H2O and solutions containing it should not be autoclaved.

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Carbonate-Bicarbonate Buffer (pH 9.2 to 10.6)

Adjustable formulation, pH 9.2–10.6 · amounts depend on target pH — use the interactive calculator

Commonly used for various immunoassay applications and for many protein and antibody conjugation procedures, including sandwich ELISA, which require experimental surface coatings. Carbonate-bicarbonate buffer is used extensively in molecular and cell biology, biochemistry, and in the medical field, where it is the most commonly used small intestine buffer. It has good buffering capacity and is easy to prepare, with excellent shelf life.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add Sodium bicarbonate of Sodium bicarbonate to the solution
3. Add Sodium carbonate (anhydrous) of Sodium carbonate (anhydrous) to the solution
4. Add distilled water until the volume is 1 L.

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TE Buffer 10X

Fixed formulation, amounts given per 1 L of final solution

TE buffer is used as a protective measure against DNA and RNA degredation, storing the two molecules and maintaining proper pH levels.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 15.759 g of Tris-Cl (desired pH) to the solution
3. Add 2.92 g of EDTA (pH 8) to the solution
4. Add distilled water until the volume is 1 L.

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Elution Buffer

Fixed formulation, amounts given per 1 L of final solution

Elution buffer is commonly used in many applications such as affinity chromatography, immunoprecipitation, protein purification, and DNA extraction to elute proteins or DNA from a ligand or membrane.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 23.38 g of Sodium chloride to the solution
3. Add distilled water until the volume is 1 L.
4. Filter the solution through a nitrocellulose filter (0.45-μM pore size) and store at room temperature.

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BAC DNA Microinjection Buffer

Fixed formulation, amounts given per 1 L of final solution

BAC injection buffer functions to stabilize BAC DNA in preparation for microinjection.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 1.211 g of Tris (pH 7.5) to the solution
3. Add 0.02922 g of EDTA to the solution
4. Add 5.844 g of Sodium chloride to the solution
5. Adjust the pH to 7.4.
6. Add distilled water until the volume is 1 L.
7. Filter the injection buffer (through a 0.2-U filter) and store at 4°C.

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EDTA Solution

Fixed formulation, amounts given per 1 L of final solution

Metal Stripping Solution is comprised of EDTA, which can act as a metal chelating agent.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 11.7 g of EDTA to the solution
3. Add distilled water until the volume is 1 L.
4. Slowly adjust the pH to 8.0 with 2 M NaOH. (Unless using the salt, EDTA often requires extended time and basic pH conditions to completely dissolve.) Store for up to 3 mo at 4°C.

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ACK Lysis Buffer

Fixed formulation, amounts given per 1 L of final solution

ACK Lysis Buffer is used to lyse red blood cells.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 8.02 g of Ammonium chloride to the solution
3. Add 1 g of Potassium bicarbonate to the solution
4. Add 0.0372 g of Disodium EDTA to the solution
5. Adjust the pH to 7.2-7.4.
6. Add distilled water until the volume is 1 L.
7. Store for up to 6 mo at room temperature.

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Paraformaldehyde Solution (8%)

Fixed formulation, amounts given per 1 L of final solution

Paraformaldehyde solution is commonly used to fix cells or tissues.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 80 g of paraformaldehyde to the solution
3. Heat to 60°C and add 2-3 drops of 1 N NaOH.
4. Add distilled water until the volume is 1 L.
5. Store this stock solution at 4°C.

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Gey's Balanced Salt Solution

Fixed formulation, amounts given per 1 L of final solution

Gey’s Balanced Salt Solution can be used for cell suspension or as washing solution.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 0.22 g of Calcium chloride dihydrate to the solution
3. Add 0.37 g of Potassium Chloride to the solution
4. Add 0.03 g of Potassium Phosphate Monobasic to the solution
5. Add 0.21 g of Magnesium Chloride Hexahydrate to the solution
6. Add 0.07 g of Magnesium Sulfate Heptahydrate to the solution
7. Add 8 g of Sodium chloride to the solution
8. Add 0.227 g of Sodium bicarbonate to the solution
9. Add 0.12 g of Sodium Phosphate Dibasic to the solution
10. Add 1 g of D-glucose to the solution
11. Add distilled water until the volume is 1 L.
12. Sterile filter and store in 500-mL aliquots at 4°C.

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Lysis Solution

Fixed formulation, amounts given per 1 L of final solution

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 146.1 g of Sodium chloride to the solution
3. Add 37.2 g of Disodium EDTA to the solution
4. Add 1.2 g of Tris base to the solution
5. Add 8 g of Sodium Hydroxide to the solution
6. Add 10 g of N-lauroylsarcosine sodium salt to the solution
7. Adjust pH to 10.
8. Add distilled water until the volume is 1 L.
9. Store at room temperature.

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H-50 Buffer

Fixed formulation, amounts given per 1 L of final solution

H-50 buffer is commonly used for washing and resuspending cells. It contains HEPES, one of the Good's zwitterionic buffers.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 4.76 g of Hepes to the solution
3. Add 3.73 g of Potassium Chloride to the solution
4. Add 0.12 g of Magnesium sulfate to the solution
5. Add 0.58 g of Sodium chloride to the solution
6. Add 0.42 g of Sodium bicarbonate to the solution
7. Add 0.156 g of Sodium phoshpate monobasic dihydrate to the solution
8. Add distilled water until the volume is 1 L.
9. Adjust the pH to 7.0 with HCl or NaOH, as appropriate. Autoclave and store cold or frozen.

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Alkaline Sodium Thiosulfate

Fixed formulation, amounts given per 1 L of final solution

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 200 g of Sodium thiosulfate to the solution
3. Add 4 g of Sodium Hydroxide to the solution
4. Add distilled water until the volume is 1 L.

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Osmium Tetroxide Solution

Fixed formulation, amounts given per 1 L of final solution

Osmium tetroxide solution is used as a fixative in electron microscopy.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 40 g of osmium tetroxide to the solution
3. Add distilled water until the volume is 1 L.
4. Make sure that the lid is tight and well sealed. Wrap in aluminum foil and stand the bottle in a larger container, sealed with a screw cap. Leave the osmium tetroxide to dissolve for 2-3 days at 4°C. Dilute 1 in 5 in double-distilled H2O before use. Osmium tetroxide solutions may be stored at -20°C for up to 6 months.

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DEAE-dextran

Fixed formulation, amounts given per 1 L of final solution

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 50 g of DEAE-dextran to the solution
3. Add distilled water until the volume is 1 L.
4. Sterilize the solution by autoclaving for 20 minutes at 15 psi (1.05 kg/cm2) on liquid cycle. Autoclaving also assists dissolution of the polymer. The molecular weight of the DEAE-dextran originally used for transfection was >2 x 106. Although this material is no longer available commercially, it is still occasionally found in chemical storerooms. The older batches of higher-molecular-weight DEAE-dextran are more efficient for transfection than the lower-molecular-weight polymers currently available.

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Trichloroacetic Acid (TCA)

Fixed formulation, amounts given per 1 L of final solution

TCA (trichloroacetic acid) is used for protein and DNA precipitation.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 1101.32 g of Trichloroacetic acid (TCA) to the solution
3. Add distilled water until the volume is 1 L.
4. The resulting solution will contain 100% (w/v) TCA.

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Bradford Stock Solution

Fixed formulation, amounts given per 1 L of final solution

1. Prepare 0 mL of distilled water in a suitable container.
2. Add 0.333 L of Ethanol (95%) to the solution
3. Add 0.667 L of Phosphoric acid (88%) to the solution
4. Add 1.16 g of Coomassie Brilliant Blue to the solution
5. Add distilled water until the volume is 1 L.

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Anesthetic (MS222) Solution

Fixed formulation, amounts given per 1 L of final solution

Anesthetic Solution (Buffered MS222) is commonly used to temporarily immobilize fish and amphibians.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 1 g of MS222 (3-aminobenzoic acid ethyl ester; Sigma-Aldrich A5040) to the solution
3. Add 0.7 g of Sodium bicarbonate to the solution
4. Add distilled water until the volume is 1 L.
5. The resulting pH should be ~7.0. Store in a foil-covered container at 4°C for up to 1 mo. Warm to room temperature before use.

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5-Fluorocytosine (5-FC) Solution

Fixed formulation, amounts given per 1 L of final solution

5-Fluorocytosine (5-FC) is a commonly used antifungal that inhibits DNA and RNA synthesis.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 10 g of 5-FC to the solution
3. Add distilled water until the volume is 1 L.
4. Vortex. Incubate for 2 h at 37°C, manually mixing the solution every 15 min until the 5-FC dissolves. Filter the solution and use it immediately. Alternatively, aliquot into sterile tubes and store at -20°C.

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SSC (Northern Blot, 20x)

Fixed formulation, amounts given per 1 L of final solution

SSC (saline sodium citrate) buffer is commonly used in Northern blotting as a hybridization or wash buffer.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 175.3 g of Sodium chloride to the solution
3. Add 77.4 g of Trisodium citrate to the solution
4. Add distilled water until the volume is 1 L.
5. Adjust the pH to 7.0 with NaOH or HCl. Autoclave the solution and store it at room temperature. It will last for 1 yr or longer.

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SSPE (Blot, 20x, pH 7.4)

Fixed formulation, amounts given per 1 L of final solution

SSPE (saline sodium phosphate EDTA) is used in hybridization and bacterial screening.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 175.3 g of Sodium chloride to the solution
3. Add 27.6 g of Sodium Phosphate Monobasic Monohydrate to the solution
4. Add 7.4 g of EDTA to the solution
5. Add distilled water until the volume is 1 L.
6. Adjust the pH to 7.4 with 10 N NaOH. Store at room temperature.

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Immunofluorescence Quenching Solution

Fixed formulation, amounts given per 1 L of final solution

Quenching solution is used in immunoflorescence and immunocytochemistry applications.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 3.75 g of Glycine to the solution
3. Add 2.67 g of Ammonium chloride to the solution
4. Add distilled water until the volume is 1 L.
5. Quenching solution can be stored at ~20°C for several weeks.

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Nuclear Fast Red Solution

Fixed formulation, amounts given per 1 L of final solution

Nuclear fast red is an anthrquinone dye used for nuclear staining.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 1 g of nuclear fast red (Kernechtrot; C.I. 60760) to the solution
3. Add 50 g of aluminum sulfate (Al2[SO4]3•18H2O) to the solution
4. Add distilled water until the volume is 1 L.
5. Heat with stirring until nearly boiling, then leave overnight to cool. There is a substantial residue of insoluble material, which must be removed by decanting and filtering. The solution keeps for ~1 yr; it may need to be filtered before each use.

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Pyronin Y (PY) Solution

Fixed formulation, amounts given per 1 L of final solution

Pyronin Y(G) (PY) is a dye used for DNA staining. PY has orange spectral emission that is easily distinguishable from FITC and 7-AAD.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 1 g of Pyronin Y(G) (PY) (Polysciences, Inc.) to the solution
3. Add distilled water until the volume is 1 L.
4. Store at 4°C protected from light.

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Mayer's Hemalum

Fixed formulation, amounts given per 1 L of final solution

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 50 g of aluminum potassium sulfate (KAl[SO4]2•12H2O) to the solution
3. Add 1 g of hematoxylin (C.I. 75290) to the solution
4. Add 0.1 g of sodium iodate (NaIO3) to the solution
5. Add 1 g of citric acid (monohydrate) to the solution
6. Add 50 g of chloral hydrate to the solution
7. A magnetic stirrer will hasten dissolving of the alum, hematoxylin, and hematein formed by iodate oxidation.
8. Add distilled water until the volume is 1 L.
9. Mayer's hemalum often keeps for 1 yr but some batches lose their potency after only a few months. The solution may be reused many times.

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Chloroquine

Fixed formulation, amounts given per 1 L of final solution

Chloroquine solution has been used to remove red blood cell-bound immunoglobin.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 52 g of Chloroquine diphosphate to the solution
3. Add distilled water until the volume is 1 L.
4. Sterilize the solution by passing it through a 0.22-µm filter; store the filtrate in foil-wrapped tubes at -20°C.

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Ethidium Bromide Solution

Fixed formulation, amounts given per 1 L of final solution

Ethidium bromide is commonly used nucleic acid stain for gel electrophoresis.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 10 g of Ethidium bromide to the solution
3. Add distilled water until the volume is 1 L.
4. Stir on a magnetic stirrer for several hours to ensure that the dye has dissolved. Wrap the container in aluminum foil or transfer the 10 mg/mL solution to a dark bottle and store at room temperature.

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Cresol Red Solution

Fixed formulation, amounts given per 1 L of final solution

Cresol red is used as a tracking dye for DNA/RNA in agarose gel electrophoresis and protein in polyacrylamide gel electrophoresis.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 4000 g of sodium salt of cresol red (Aldrich) to the solution
3. Add distilled water until the volume is 1 L.
4. Store the 10 mM solution at room temperature.

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Bath Solution (Ca2+/Mg2+ Free)

Fixed formulation, amounts given per 1 L of final solution

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 7.854 g of Potassium hydroxide to the solution
3. Add 4.766 g of Hepes to the solution
4. Add 0.1491 g of Potassium Chloride to the solution
5. Add 1.902 g of EGTA to the solution
6. Add 13.314 g of Methanesulfonic acid to the solution
7. Add distilled water until the volume is 1 L.
8. Use methanesulfonic acid to adjust the pH to 7.1-7.2. Store for up to 3 years at room temperature (22°C-24°C).

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Modified Barth's Saline (MBS)

Fixed formulation, amounts given per 1 L of final solution

Modified Barth's Saline (MBS) is commonly used as culture media for Xenopus oocytes

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 11.92 g of Hepes to the solution
3. Add 51.43 g of Sodium chloride to the solution
4. Add 0.75 g of Potassium Chloride to the solution
5. Add 2.1 g of Sodium bicarbonate to the solution
6. Add 2.46 g of Magnesium Sulfate Heptahydrate to the solution
7. Adjust the pH of 10x MBS salt solution to 7.8 with 10 M NaOH, and sterilize by autoclaving. Store indefinitely at room temperature.
8. Add distilled water until the volume is 1 L.
9. To prepare 1x MBS, combine 100 mL of 10x MBS salt solution and 7 mL of 0.1 M CaCl2 with 893 mL of autoclaved deionized H2O. Add antibiotics. Store for up to 1-2 mo at 16°C-18°C (the same temperature used for oocyte incubation). Check before use.

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Calcium Chloride Stock Solution

Fixed formulation, amounts given per 1 L of final solution

Calcium chloride is a common calcium salt used in a variety of applications.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 58 g of Calcium Chloride to the solution
3. Add distilled water until the volume is 1 L.
4. Measure the osmolarity using an osmometer: if below 1000 mmol/kg, add more CaCl2; if above 1000, add H2O. (The osmolarity of 1000 mmol/kg is ~0.383 M. Preparation of this calibrated solution is necessary because calcium chloride is hygroscopic, so is not reliably measured by weight. Slight errors in calcium concentration have an outsized effect on synaptic physiology.) Store the solution at 4?C. Solution stored under these conditions appears to be stable for several months.

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Yeast Two Hybrid (Y2H) Media, Amino Acid Dropout Mixes

Fixed formulation, amounts given per 1 L of final solution

Amino acid dropout mixtures contain amino acid supplements necessary for the growth of yeast. This amino acid dropout mixture is suitable for use with yeast-two hybrid systems.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 0.01 g of Adenine to the solution
3. Add 0.05 g of L-Arginine to the solution
4. Add 0.08 g of L-Aspartic acid to the solution
5. Add 0.02 g of L-Histidine to the solution
6. Add 0.05 g of L-Isoleucine to the solution
7. Add 0.1 g of L-Leucine to the solution
8. Add 0.05 g of L-Lysine to the solution
9. Add 0.02 g of L-Methionine to the solution
10. Add 0.05 g of L-Phenylalanine to the solution
11. Add 0.1 g of L-Threonine to the solution
12. Add 0.05 g of L-Tryptophan to the solution
13. Add 0.05 g of L-Tyrosine to the solution
14. Add 0.14 g of L-Valine to the solution
15. Add 0.02 g of Uracil to the solution
16. Add distilled water until the volume is 1 L.
17. Combine the appropriate ingredients, and mix in a sealed container. Omit the desired dropout component. Haploid yeast requires a single dropout (i.e., -Trp=-T or -Leu=-L); diploid yeast requires double dropouts (i.e., -Leu -Trp=-LT); and selecting for interactions requires triple dropouts (i.e., -Leu-Trp -His=-LTH, plus 3-AT).

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Digestion Medium

Fixed formulation, amounts given per 1 L of final solution

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 23.4 g of Sodium chloride to the solution
3. Add 26.4 g of Magnesium Chloride Hexahydrate to the solution
4. Add 5.4 g of Magnesium Sulfate Heptahydrate to the solution
5. Add 11.9 g of Potassium Chloride to the solution
6. Add 0.3 g of Calcium Chloride to the solution
7. Add 1.95 g of 2-(N-morpholino)ethanesulfonic acid to the solution
8. Add distilled water until the volume is 1 L.
9. Adjust the pH to 6.5. Filter through a 0.2-µm filter and store at 4°C. Just before use, add 1% (w/v) cellulose (Sigma) and 3 U/mL alginate lyase (Boyen et al. 1990).

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Chelation Medium

Fixed formulation, amounts given per 1 L of final solution

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 40.9 g of Sodium chloride to the solution
3. Add 6.1 g of Magnesium Chloride Hexahydrate to the solution
4. Add 7.39 g of Magnesium Sulfate Heptahydrate to the solution
5. Add 1.49 g of Potassium Chloride to the solution
6. Add 7.6 g of EGTA to the solution
7. Add distilled water until the volume is 1 L.
8. Adjust pH to 5.5 using HCl. Autoclave or filter through a 0.2-µm membrane. Store at 4°C.

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Penicillin/Streptomycin/Chloramphenicol Antibiotic Mix

Fixed formulation, amounts given per 1 L of final solution

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 10 g of Penicillin G to the solution
3. Add 5 g of Streptomycin to the solution
4. Add 1 g of Chloramphenicol to the solution
5. Add distilled water until the volume is 1 L.
6. Filter through a 0.22-µm membrane and store in 5-mL aliquots at ~20°C. For use, add 5 mL of the antibiotic mix to 1 L of Provasoli-enriched seawater (PES).

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Embryonic Medium

Fixed formulation, amounts given per 1 L of final solution

Embryonic medium is used for culturing embryos.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 0.87 g of Sodium chloride to the solution
3. Add 0.04 g of Potassium Chloride to the solution
4. Add 0.15 g of Calcium chloride dihydrate to the solution
5. Add 0.25 g of Magnesium Sulfate Heptahydrate to the solution
6. Add 0.02 g of Potassium Phosphate Monobasic to the solution
7. Add 0.02 g of Sodium phosphate dibaic dodecahydrate to the solution
8. Add 0.06 g of Sodium bicarbonate to the solution
9. Add distilled water until the volume is 1 L.
10. Adjust the pH to 7.0-7.5.

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Carbenicillin Solution

Fixed formulation, amounts given per 1 L of final solution

Carbenicillin is a bactericidal antibiotic that has beta-lactamase resistant activity.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 50 g of disodium carbenicillin to the solution
3. Add distilled water until the volume is 1 L.
4. Filter-sterilize using a 0.22-µm filter. Store in 1-mL aliquots at ~20°C.

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M9 Minimal Salts

Fixed formulation, amounts given per 1 L of final solution

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 64 g of Sodium Phosphate Dibasic Heptahydrate to the solution
3. Add 15 g of Potassium Phosphate Monobasic to the solution
4. Add 2.5 g of Sodium chloride to the solution
5. Add 5 g of Ammonium chloride to the solution
6. Add distilled water until the volume is 1 L.
7. Divide the salt solution into 200-ml aliquots and sterilize by autoclaving for 15 minutes at 15 psi (1.05 kg/cm 2) on liquid cycle.

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IPTG Solution

Fixed formulation, amounts given per 1 L of final solution

IPTG (isopropyl-beta-D-thiogalactopyranoside) is a synthetic lactose analog.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 200 g of IPTG to the solution
3. Add distilled water until the volume is 1 L.
4. Sterilize by passing it through a 0.22-µm disposable filter. Dispense the solution into 1-ml aliquots and store them at -20°C.

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SDS-PAGE SDS Running Buffer (10x)

Fixed formulation, amounts given per 1 L of final solution

In SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis), SDS Running Buffer is used as the electrophoresis buffer during stacking and resolution.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 30.3 g of Tris base to the solution
3. Add 144.4 g of Glycine to the solution
4. Add 10 g of SDS to the solution
5. Add distilled water until the volume is 1 L.

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Electrophoretic Transfer Buffer (20x)

Fixed formulation, amounts given per 1 L of final solution

Electrophoretic Transfer Buffers are used in SDS-PAGE to transfer proteins from a gel to a nitrocellulose membrane.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 35.29 g of Trisodium citrate to the solution
3. Add 22.71 g of Sodium phosphate, dibasic (Na2HPO4) to the solution
4. Add distilled water until the volume is 1 L.
5. Adjust the pH to 8.3, autoclave, and store for up to 1-2 yr at room temperature.

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Tris-Glycine Transfer Buffer (20x)

Fixed formulation, amounts given per 1 L of final solution

Tris-Glycine Transfer Buffer is used as a transfer buffer in Western blotting.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 24.2 g of Tris base to the solution
3. Add 150.1 g of Glycine to the solution
4. Add distilled water until the volume is 1 L.
5. pH adjustment is not necessary (it will be ~8.8). Store at room temperature.

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Sodium Carbonate Transfer Buffer (40x, pH 9.5)

Fixed formulation, amounts given per 1 L of final solution

Sodium Carbonate Transfer Buffer is used to transfer proteins following SDS-PAGE.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 21.1 g of Sodium bicarbonate to the solution
3. Add 18.35 g of Sodium carbonate to the solution
4. Adjust the pH to 9.5 with 10 N NaOH.
5. Add distilled water until the volume is 1 L.
6. Store at room temperature.

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Acrylamide Stock Solution

Fixed formulation, amounts given per 1 L of final solution

Acrylamide is used along with bisacrylamide to form gels used in SDS-PAGE. Acrylamide forms polyacrylamide polymers, while bisacrylamide crosslinks between polyacrylamide chains.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 389.6 g of acrylamide to the solution
3. Add 10.4 g of bisacrylamide to the solution
4. Add distilled water until the volume is 1 L.
5. Filter the stock solution using Whatman filter paper and store at 4 °C. Prepare fresh acrylamide stock solution every few weeks.

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Imidazole-SDS Solution

Fixed formulation, amounts given per 1 L of final solution

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 13.6 g of Imidazole to the solution
3. Add 1 g of SDS to the solution
4. Add distilled water until the volume is 1 L.

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Electrode Buffer

Fixed formulation, amounts given per 1 L of final solution

Electrode buffer has been used in studies involving polymer solar cells.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 11.6 g of Tris base to the solution
3. Add 37.5 g of Glycine to the solution
4. Add 3.98 g of SDS to the solution
5. Add distilled water until the volume is 1 L.

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Tris/EACA Buffer

Fixed formulation, amounts given per 1 L of final solution

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 1.31 g of ε-aminocaproic acid (EACA) to the solution
3. Add 10.9 g of Tris base to the solution
4. Add distilled water until the volume is 1 L.

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PBS (Phosphate Buffered Saline) (1X, pH 7.4)

Fixed formulation, amounts given per 1 L of final solution

PBS is an isotonic buffer frequently used in biological applications, such as washing cells, transportation of tissues, and dilutions. PBS closely mimics the pH, osmolarity, and ion concentrations of the human body. Since it is nontoxic to cells, it is extensively used for cell container rinsing and other preparations that might leave a residue. It is simple to prepare and has good shelf life, but will precipitate in the presence of zinc ions.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 8 g of Sodium chloride to the solution
3. Add 0.2 g of Potassium Chloride to the solution
4. Add 1.44 g of Sodium Phosphate Dibasic to the solution
5. Add 0.245 g of Potassium Phosphate Monobasic to the solution
6. Adjust solution to desired pH (typically pH ≈ 7.4).
7. Add distilled water until the volume is 1 L.

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HBSS (Hank's Balanced Salt Solution)

Fixed formulation, amounts given per 1 L of final solution

Maintains physiological pH and osmotic balance for cells.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 8 g of Sodium chloride to the solution
3. Add 0.4 g of Potassium Chloride to the solution
4. Add 0.14 g of Calcium Chloride to the solution
5. Add 0.1 g of Magesium Sulfate Heptahydrate to the solution
6. Add 0.1 g of Magnesium Chloride Hexahydrate to the solution
7. Add 0.06 g of Sodium Phosphate Dibasic Dihydrate to the solution
8. Add 0.06 g of Potassium Phosphate Monobasic to the solution
9. Add 1 g of D-Glucose (Dextrose) to the solution
10. Add 0.35 g of Sodium bicarbonate to the solution
11. Add distilled water until the volume is 1 L.

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HHBS (Hank's Buffer with Hepes)

Fixed formulation, amounts given per 1 L of final solution

HHBS is a general purpose buffer that combines the stability of Hank's buffer with the good buffering capacity of HEPES buffering solution (one of Good's buffers). HHBS is used extensively for cell culture, but is sensitive to light.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 0.14 g of Calcium Chloride to the solution
3. Add 0.1 g of Magnesium Chloride Hexahydrate to the solution
4. Add 0.1 g of Magesium Sulfate Heptahydrate to the solution
5. Add 0.4 g of Potassium Chloride to the solution
6. Add 0.06 g of Potassium Phosphate Monobasic to the solution
7. Add 0.35 g of Sodium bicarbonate to the solution
8. Add 8 g of Sodium chloride to the solution
9. Add 0.048 g of Sodium Phosphate Dibasic to the solution
10. Add 1 g of D-Glucose (Dextrose) to the solution
11. Add 4.76 g of Hepes to the solution
12. Add distilled water until the volume is 1 L.

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HEPES Buffer (1 M, 7.5 pH)

Fixed formulation, amounts given per 1 L of final solution

HEPES buffer is one of the Good's zwitterionic buffer with a pH range of 6.8-8.2. It is commonly used in cell culture medium as its pH is well maintained with changes in carbon dioxide concentration.

1. Prepare 800 mL of dH2O in a suitable container.
2. Add 238.3 g of Hepes to the solution
3. Adjust solution to desired pH by 10N NaOH.
4. Add dH2O until the volume is 1 L.

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PBS with Ca2+and Mg2+ (D-PBS)

Fixed formulation, amounts given per 1 L of final solution

DPBS (Dulbecco's phosphate-buffered saline) with Ca2+ and Mg2+ is widely used for reagent preparation and as transport media, as well as washing and dilution.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 0.1 g of Calcium Chloride to the solution
3. Add 0.2 g of Potassium Chloride to the solution
4. Add 0.1 g of Magnesium Chloride Hexahydrate to the solution
5. Add 8 g of Sodium chloride to the solution
6. Add 2.16 g of Sodium Phosphate Dibasic to the solution
7. Add distilled water until the volume is 1 L.
8. Filter-sterilize. Store at 25°C for up to 12 mo.

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Extracellular Saline (1x, pH 7.3)

Fixed formulation, amounts given per 1 L of final solution

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 8.182 g of Sodium chloride to the solution
3. Add 0.402 g of Potassium Chloride to the solution
4. Add 0.294 g of Calcium Chloride to the solution
5. Add 1.204 g of Magnesium sulfate to the solution
6. Add 1.191 g of Hepes to the solution
7. Add 1.802 g of Glucose to the solution
8. Add distilled water until the volume is 1 L.

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Tyrode's Solution, Acidic

Fixed formulation, amounts given per 1 L of final solution

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 8 g of Sodium chloride to the solution
3. Add 0.2 g of Potassium Chloride to the solution
4. Add 0.24 g of Calcium chloride dihydrate to the solution
5. Add 0.1 g of Magnesium Chloride Hexahydrate to the solution
6. Add 1 g of Glucose to the solution
7. Add 4 g of Polyvinylpyrrolidone (PVP) to the solution
8. Add distilled water until the volume is 1 L.
9. Prepare at room temperature and adjust to pH 2.5 with Analar HCl (BDH). The PVP is added to increase viscosity and reduce embryo stickiness. Filter-sterilize and store in aliquots at -20°C.

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Sodium Iodide/Sodium Sulfite Solution

Fixed formulation, amounts given per 1 L of final solution

Sodium Iodide/Sodium Sulfite Solution can be used as a chaotropic agent in DNA recovery.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 1 g of Sodium sulfite (Na2SO3) to the solution
3. Add 675 g of Sodium iodide to the solution
4. Add distilled water until the volume is 1 L.
5. Filter the solution through Whatman paper and use immediately.

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Dithiothreitol (DTT) (1 M)

Fixed formulation, amounts given per 1 L of final solution

DTT (Dithiothreitol) is a commonly used reducing agent.

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 150 g of DTT (DL-dithiothreitol, anhydrous m.w. = 154.25) to the solution
3. Add distilled water until the volume is 1 L.
4. Dispense into 1-mL aliquots, and store them in the dark (wrapped in aluminum foil) at -20°C (indefinitely). Do not autoclave DTT or solutions containing it.

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