常用试剂配制－3

Chrom alum gelatin (Subbing solution)

Dissolve 5.0 gram of gelatin in 1 liter of boiling water. Cool and add 0.5 grams of potassium chrome alum (CrK(SO ) ・ 12H O). Store in refrigerator.
To use, dip clean slides into the solution and dry in vertical position in a dust free location.

 

Citric acid (H C H O ・ H O MW 210.14)

0.1 M

Dissolve 21.01 grams of citric acid to a final volume of 1 liter.

 

Citrate buffer (Sodium phosphate-Citrate buffer)

0.001 M
PH 4.8	Add 493 ml of 0.2 M Na HPO to 507 ml of 0.1 M citric acid.
PH 3.6	Add 322 ml of 0.2 M Na HPO to 678 ml of 0.1 M citric acid.
PH 4.2	Add 414 ml of 0.2 M Na HPO to 586 ml of 0.1 M citric acid.
PH 5.4	Add 557.5 ml of 0.2 M Na HPO to 442.6 ml of 0.1 M citric acid.
PH 6.0	Add 631.5 ml of 0.2 M Na HPO to 368.5 ml of 0.1 M citric acid.
PH 6.6	Add 727.5 ml of 0.2 M Na HPO to 272.5 ml of 0.1 M citric acid.
PH 7.2	Add 869.5 ml of 0.2 M Na HPO to 130.5 ml of 0.1 M citric acid.
PH 7.8	Add 957.5 ml of 0.2 M Na HPO to 42.5 ml of 0.1 M citric acid.

 

Cobaltous Nitrate (MW 182.96)

2% (w/v)

Dissolve 2.0 grams of cobaltous nitrate (hexahydrate is very soluble) to a final volume of 100 ml with water. Keep well closed in a cool place.

 

Colcemid

10 µ g/ml

Dissolve 10 g of colcemid per ml of saline or culture medium.

 

Coomasie blue (Coomasie Brilliant Blue R250)

0.25% (w/v)0.001 M

Dissolve 2.50 grams of Coomasie Brilliant Blue R250 to a final volume of 1 liter with 20% (w/v) trichloroacetic acid (TCA). Some investigators use a 0.25% solution of Coomasie Blue in methanol-water-glacial acetic acid (5-5-1).

 

Copper sulfate (CuSO 5H O MW 249.68)

0.5% (w/v)

Dissolve 0.13 grams of Alcian Blue 8GX (Sigma # A-2899) in 100 ml of water.

 

0.5% (w/v)

Dissolve 0.5 grams of copper sulfate to a final volume of 100 ml with water.

 

Copper tartrate/carbonate (CTC)

Dissolve 0.5 grams of copper sulfate and 1.0 gram of potassium sodium tartrate to a final volume of 100 ml with water. Combine 1.0 ml of this solution with 50 ml of 2% Na CO in 0.1 N NaOH. Must be made fresh, prior to use. Stock solutions are stable.

 

Crystal Violet

Dissolve 0.1 grams of crystal violet and 0.25 ml of glacial acetic acid to a final volume of 100 ml with water.

 

DCMU (3-(3,4-Dichlorophenyl)-1,1-Dimethylurea MW 233.1)

1 x 10 M 0.001 M	Dissolve 2.3 mg DCMU to a final volume of 100 ml with water or buffer.
5 x 10 M0.001 M	Dilute the 1 x 10- M solution 1/200 prior to use.

 

Dichlorophenolindophenol (DCPIP MW 290.1)

0.0025 M	Dissolve 73 mg of DCPIP to a final volume of 100 ml with water or buffer.
0.0001 M	Dissolve 2.9 mg of DCPIP to a final volume of 100 ml with water or buffer.

 

Dinitrophenol (DNP MW 184.11)

18.4 mg%

Dissolve 18.4 mg of 2,4-dinitrophenol to a final volume of 100 ml with water or buffer.

 

Dische diphenylamine reagent

Dissolve 500 mg of diphenylamine in 49 ml of glacial acetic acid. Add 1.0 ml of concentrated HCl.

 

Dithiothreitol (Cleland's Reagent MW 154.3)

0.01 M

Dissolve 154 mg of dithiothreitol to a final volume of 100 ml with water or buffer. Dithiothreitol is available from Sigma Chemical Co., St. Louis, Cat # D0632. Dithioerythritol may be substituted.

 

DOPA (3-(3,4-Dihydroxyphenyl)-L-alanine MW 197.19)

8 mM	Dissolve 158 mg of L-DOPA to a final volume of 100 ml with water or buffer. Note that the maximum solubility of DOPA in water is 165 mg/100 ml (8.3 mM).

 

EDTA (Ethylenediaminetetraacetic acid MW 292.24)

1 M

Dissolve 292.24 grams of EDTA, free acid to a final volume of 1 liter. If the more soluble disodium salt of EDTA is used, adjust the weight accordingly. The pH can be adjusted with acetic acid or NaOH. For corresponding concentration dilutions, multiply the weight in grams by the desired molarity. For exam ple, for 10 mM EDTA, multiply 292.24 X 0.010 to obtain 2.92 grams of EDTA per liter.

 

EGTA (Ethylene Glycol-bis( aminoethyl Ether) N,N,N',N'-Tetraacetic Acid MW 380.4)

1 mM	issolve 380 mg of EGTA to a final volume of 1 liter with water or buffer.

 

Eosin

0.5% (w/v)

Dissolve 0.5 grams of Eosin Y in 100 ml of water.

 

Ethanol (C H OH MW 46.07)

Density = 0.7893 gm/ml

50-95% (v/v)

Since 95% ethyl alcohol is less expensive and easier to store than absolute, these dilutions should be made with 95% ethyl alcohol. Unless otherwise stated, denatured alcohol works as well as the more expensive non-denatured. A simple way to make the % so lution is to use the appropriate amount of 95% ethanol and dilute to 950 ml instead of 1 liter. For example, to make a 50% (v/v) solution, measure out 500 ml of 95% ethyl alcohol and dilute to a final volume of 950 ml with water. For a 70% solution, meas ure 700 ml of ethyl alcohol and dilute to 950 ml with water. Absolute ethanol should be used directly as 100% ethanol. It is important for histology that this be truly 100%. Since it is hydroscopic (it absorbs water from the air), do not assume it is abso lute unless it is sealed or treated to ensure no water. To test, add a drop to a sample of xylol. If any cloudiness occurs, the alcohol is not absolute.

8.5 M

The ethanol can be weighed (391.6 grams of absolute, 412.2 grams of 95% (v/v) or measured volumetrically by using the density. That is, 391.6 grams  0.7893 grams/ml or 496.1 ml of absolute ethanol. Using 95%, 412.2 grams  0.7893 grams /ml or 522.2 ml. Weigh or measure the appropriate amount and dilute to a final volume of 1 liter with water.

 

Ethanol-acetic acid fixative for histology (3:1)

To 75 ml of absolute alcohol, add 25 ml of glacial acetic acid. Must be made fresh, just prior to use.

 

Fetal Calf Serum (FCS)

While it is possible to prepare your own serum from whole blood, it is easier (and safer) to purchase FCS from a reputable supplier. Commercial sources are free of mycoplasma, pre-sterilized and controlled for the presence of antibodies. There are a n umber of serum substitutes available on the market and these may be less expensive when storage is considered. Suppliers include Gibco, Flow Laboratories, KC Biological and Sigma Chemical Co.  

 

Folin-Ciocalteu Reagent

This is usually purchased premixed, since it is difficult to make. Also known as 2N Folin-phenol reagent.

 

Giemsa stain

Prepare a stock solution by dissolving 3.8 grams of giemsa powder in 25 ml of glycerin. Heat gently with stirring for about 2 hours at 60 ° C. Cool and add 75 ml of methanol (neutral, acetone free).
For a working solution, dilute the stock solution 1/10 with water before use.
For chromosome banding, combine 5.0 ml of stock Giemsa, 3.0 ml of absolute methanol, 3.0 ml of 0.1 M citric acid and 89 ml of distilled water. Adjust the pH of the solution to 6.6 with Na HPO .

 

Glucose (MW 180.16)

10% (v/v)

Dissolve 10 grams of D-glucose (dextrose) in a final volume of 100 ml with water or buffer.

 

Glutaraldehyde (GTA)

5 %	GTA is usually supplied as a 25% or 50% (w/v) solution. It is used for electron microscope fixation as a 5% solution in a buffer. For routine use, add 20 ml of 25% GTA to 80 ml of 0.2 M sodium cacodylate buffer, pH 7.4.

 

Glycerol (MW 92.09)

10% (v/v)	To 10 ml of glycerol (glycerine) add enough water to make a final volume of 100 ml.
8 M	Weigh 73.67 grams of glycerol and add to a final volume of 100 ml. Alternatively, measure 499.1 ml of glycerol and make to a final volume of 1 liter (the density of glycerol at room temperature is 1.476) with water or buffer. For 8 M glycerol in MT buffer, make a 2X MT buffer for use as the diluent.

Chrom alum gelatin (Subbing solution)

Dissolve 5.0 gram of gelatin in 1 liter of boiling water. Cool and add 0.5 grams of potassium chrome alum (CrK(SO ) ・ 12H O). Store in refrigerator.
To use, dip clean slides into the solution and dry in vertical position in a dust free location.

 

Citric acid (H C H O ・ H O MW 210.14)

0.1 M

Dissolve 21.01 grams of citric acid to a final volume of 1 liter.

 

Citrate buffer (Sodium phosphate-Citrate buffer)

0.001 M
PH 4.8	Add 493 ml of 0.2 M Na HPO to 507 ml of 0.1 M citric acid.
PH 3.6	Add 322 ml of 0.2 M Na HPO to 678 ml of 0.1 M citric acid.
PH 4.2	Add 414 ml of 0.2 M Na HPO to 586 ml of 0.1 M citric acid.
PH 5.4	Add 557.5 ml of 0.2 M Na HPO to 442.6 ml of 0.1 M citric acid.
PH 6.0	Add 631.5 ml of 0.2 M Na HPO to 368.5 ml of 0.1 M citric acid.
PH 6.6	Add 727.5 ml of 0.2 M Na HPO to 272.5 ml of 0.1 M citric acid.
PH 7.2	Add 869.5 ml of 0.2 M Na HPO to 130.5 ml of 0.1 M citric acid.
PH 7.8	Add 957.5 ml of 0.2 M Na HPO to 42.5 ml of 0.1 M citric acid.

 

Cobaltous Nitrate (MW 182.96)

2% (w/v)

Dissolve 2.0 grams of cobaltous nitrate (hexahydrate is very soluble) to a final volume of 100 ml with water. Keep well closed in a cool place.

 

Colcemid

10 µ g/ml

Dissolve 10 g of colcemid per ml of saline or culture medium.

 

Coomasie blue (Coomasie Brilliant Blue R250)

0.25% (w/v)0.001 M

Dissolve 2.50 grams of Coomasie Brilliant Blue R250 to a final volume of 1 liter with 20% (w/v) trichloroacetic acid (TCA). Some investigators use a 0.25% solution of Coomasie Blue in methanol-water-glacial acetic acid (5-5-1).

 

Copper sulfate (CuSO 5H O MW 249.68)

0.5% (w/v)

Dissolve 0.13 grams of Alcian Blue 8GX (Sigma # A-2899) in 100 ml of water.

 

0.5% (w/v)

Dissolve 0.5 grams of copper sulfate to a final volume of 100 ml with water.

 

Copper tartrate/carbonate (CTC)

Dissolve 0.5 grams of copper sulfate and 1.0 gram of potassium sodium tartrate to a final volume of 100 ml with water. Combine 1.0 ml of this solution with 50 ml of 2% Na CO in 0.1 N NaOH. Must be made fresh, prior to use. Stock solutions are stable.

 

Crystal Violet

Dissolve 0.1 grams of crystal violet and 0.25 ml of glacial acetic acid to a final volume of 100 ml with water.

 

DCMU (3-(3,4-Dichlorophenyl)-1,1-Dimethylurea MW 233.1)

1 x 10 M 0.001 M	Dissolve 2.3 mg DCMU to a final volume of 100 ml with water or buffer.
5 x 10 M0.001 M	Dilute the 1 x 10- M solution 1/200 prior to use.

 

Dichlorophenolindophenol (DCPIP MW 290.1)

0.0025 M	Dissolve 73 mg of DCPIP to a final volume of 100 ml with water or buffer.
0.0001 M	Dissolve 2.9 mg of DCPIP to a final volume of 100 ml with water or buffer.

 

Dinitrophenol (DNP MW 184.11)

18.4 mg%

Dissolve 18.4 mg of 2,4-dinitrophenol to a final volume of 100 ml with water or buffer.

 

Dische diphenylamine reagent

Dissolve 500 mg of diphenylamine in 49 ml of glacial acetic acid. Add 1.0 ml of concentrated HCl.

 

Dithiothreitol (Cleland's Reagent MW 154.3)

0.01 M

Dissolve 154 mg of dithiothreitol to a final volume of 100 ml with water or buffer. Dithiothreitol is available from Sigma Chemical Co., St. Louis, Cat # D0632. Dithioerythritol may be substituted.

 

DOPA (3-(3,4-Dihydroxyphenyl)-L-alanine MW 197.19)

8 mM	Dissolve 158 mg of L-DOPA to a final volume of 100 ml with water or buffer. Note that the maximum solubility of DOPA in water is 165 mg/100 ml (8.3 mM).

 

EDTA (Ethylenediaminetetraacetic acid MW 292.24)

1 M

Dissolve 292.24 grams of EDTA, free acid to a final volume of 1 liter. If the more soluble disodium salt of EDTA is used, adjust the weight accordingly. The pH can be adjusted with acetic acid or NaOH. For corresponding concentration dilutions, multiply the weight in grams by the desired molarity. For exam ple, for 10 mM EDTA, multiply 292.24 X 0.010 to obtain 2.92 grams of EDTA per liter.

 

EGTA (Ethylene Glycol-bis( aminoethyl Ether) N,N,N',N'-Tetraacetic Acid MW 380.4)

1 mM	issolve 380 mg of EGTA to a final volume of 1 liter with water or buffer.

 

Eosin

0.5% (w/v)

Dissolve 0.5 grams of Eosin Y in 100 ml of water.

 

Ethanol (C H OH MW 46.07)

Density = 0.7893 gm/ml

50-95% (v/v)

Since 95% ethyl alcohol is less expensive and easier to store than absolute, these dilutions should be made with 95% ethyl alcohol. Unless otherwise stated, denatured alcohol works as well as the more expensive non-denatured. A simple way to make the % so lution is to use the appropriate amount of 95% ethanol and dilute to 950 ml instead of 1 liter. For example, to make a 50% (v/v) solution, measure out 500 ml of 95% ethyl alcohol and dilute to a final volume of 950 ml with water. For a 70% solution, meas ure 700 ml of ethyl alcohol and dilute to 950 ml with water. Absolute ethanol should be used directly as 100% ethanol. It is important for histology that this be truly 100%. Since it is hydroscopic (it absorbs water from the air), do not assume it is abso lute unless it is sealed or treated to ensure no water. To test, add a drop to a sample of xylol. If any cloudiness occurs, the alcohol is not absolute.

8.5 M

The ethanol can be weighed (391.6 grams of absolute, 412.2 grams of 95% (v/v) or measured volumetrically by using the density. That is, 391.6 grams  0.7893 grams/ml or 496.1 ml of absolute ethanol. Using 95%, 412.2 grams  0.7893 grams /ml or 522.2 ml. Weigh or measure the appropriate amount and dilute to a final volume of 1 liter with water.

 

Ethanol-acetic acid fixative for histology (3:1)

To 75 ml of absolute alcohol, add 25 ml of glacial acetic acid. Must be made fresh, just prior to use.

 

Fetal Calf Serum (FCS)

While it is possible to prepare your own serum from whole blood, it is easier (and safer) to purchase FCS from a reputable supplier. Commercial sources are free of mycoplasma, pre-sterilized and controlled for the presence of antibodies. There are a n umber of serum substitutes available on the market and these may be less expensive when storage is considered. Suppliers include Gibco, Flow Laboratories, KC Biological and Sigma Chemical Co.  

 

Folin-Ciocalteu Reagent

This is usually purchased premixed, since it is difficult to make. Also known as 2N Folin-phenol reagent.

 

Giemsa stain

Prepare a stock solution by dissolving 3.8 grams of giemsa powder in 25 ml of glycerin. Heat gently with stirring for about 2 hours at 60 ° C. Cool and add 75 ml of methanol (neutral, acetone free).
For a working solution, dilute the stock solution 1/10 with water before use.
For chromosome banding, combine 5.0 ml of stock Giemsa, 3.0 ml of absolute methanol, 3.0 ml of 0.1 M citric acid and 89 ml of distilled water. Adjust the pH of the solution to 6.6 with Na HPO .

 

Glucose (MW 180.16)

10% (v/v)

Dissolve 10 grams of D-glucose (dextrose) in a final volume of 100 ml with water or buffer.

 

Glutaraldehyde (GTA)

5 %	GTA is usually supplied as a 25% or 50% (w/v) solution. It is used for electron microscope fixation as a 5% solution in a buffer. For routine use, add 20 ml of 25% GTA to 80 ml of 0.2 M sodium cacodylate buffer, pH 7.4.

 

Glycerol (MW 92.09)

10% (v/v)	To 10 ml of glycerol (glycerine) add enough water to make a final volume of 100 ml.
8 M	Weigh 73.67 grams of glycerol and add to a final volume of 100 ml. Alternatively, measure 499.1 ml of glycerol and make to a final volume of 1 liter (the density of glycerol at room temperature is 1.476) with water or buffer. For 8 M glycerol in MT buffer, make a 2X MT buffer for use as the diluent.

Chrom alum gelatin (Subbing solution)

Dissolve 5.0 gram of gelatin in 1 liter of boiling water. Cool and add 0.5 grams of potassium chrome alum (CrK(SO ) ・ 12H O). Store in refrigerator.
To use, dip clean slides into the solution and dry in vertical position in a dust free location.

 

Citric acid (H C H O ・ H O MW 210.14)

0.1 M

Dissolve 21.01 grams of citric acid to a final volume of 1 liter.

 

Citrate buffer (Sodium phosphate-Citrate buffer)

0.001 M
PH 4.8	Add 493 ml of 0.2 M Na HPO to 507 ml of 0.1 M citric acid.
PH 3.6	Add 322 ml of 0.2 M Na HPO to 678 ml of 0.1 M citric acid.
PH 4.2	Add 414 ml of 0.2 M Na HPO to 586 ml of 0.1 M citric acid.
PH 5.4	Add 557.5 ml of 0.2 M Na HPO to 442.6 ml of 0.1 M citric acid.
PH 6.0	Add 631.5 ml of 0.2 M Na HPO to 368.5 ml of 0.1 M citric acid.
PH 6.6	Add 727.5 ml of 0.2 M Na HPO to 272.5 ml of 0.1 M citric acid.
PH 7.2	Add 869.5 ml of 0.2 M Na HPO to 130.5 ml of 0.1 M citric acid.
PH 7.8	Add 957.5 ml of 0.2 M Na HPO to 42.5 ml of 0.1 M citric acid.

 

Cobaltous Nitrate (MW 182.96)

2% (w/v)

Dissolve 2.0 grams of cobaltous nitrate (hexahydrate is very soluble) to a final volume of 100 ml with water. Keep well closed in a cool place.

 

Colcemid

10 µ g/ml

Dissolve 10 g of colcemid per ml of saline or culture medium.

 

Coomasie blue (Coomasie Brilliant Blue R250)

0.25% (w/v)0.001 M

Dissolve 2.50 grams of Coomasie Brilliant Blue R250 to a final volume of 1 liter with 20% (w/v) trichloroacetic acid (TCA). Some investigators use a 0.25% solution of Coomasie Blue in methanol-water-glacial acetic acid (5-5-1).

 

Copper sulfate (CuSO 5H O MW 249.68)

0.5% (w/v)

Dissolve 0.13 grams of Alcian Blue 8GX (Sigma # A-2899) in 100 ml of water.

 

0.5% (w/v)

Dissolve 0.5 grams of copper sulfate to a final volume of 100 ml with water.

 

Copper tartrate/carbonate (CTC)

Dissolve 0.5 grams of copper sulfate and 1.0 gram of potassium sodium tartrate to a final volume of 100 ml with water. Combine 1.0 ml of this solution with 50 ml of 2% Na CO in 0.1 N NaOH. Must be made fresh, prior to use. Stock solutions are stable.

 

Crystal Violet

Dissolve 0.1 grams of crystal violet and 0.25 ml of glacial acetic acid to a final volume of 100 ml with water.

 

DCMU (3-(3,4-Dichlorophenyl)-1,1-Dimethylurea MW 233.1)

1 x 10 M 0.001 M	Dissolve 2.3 mg DCMU to a final volume of 100 ml with water or buffer.
5 x 10 M0.001 M	Dilute the 1 x 10- M solution 1/200 prior to use.

 

Dichlorophenolindophenol (DCPIP MW 290.1)

0.0025 M	Dissolve 73 mg of DCPIP to a final volume of 100 ml with water or buffer.
0.0001 M	Dissolve 2.9 mg of DCPIP to a final volume of 100 ml with water or buffer.

 

Dinitrophenol (DNP MW 184.11)

18.4 mg%

Dissolve 18.4 mg of 2,4-dinitrophenol to a final volume of 100 ml with water or buffer.

 

Dische diphenylamine reagent

Dissolve 500 mg of diphenylamine in 49 ml of glacial acetic acid. Add 1.0 ml of concentrated HCl.

 

Dithiothreitol (Cleland's Reagent MW 154.3)

0.01 M

Dissolve 154 mg of dithiothreitol to a final volume of 100 ml with water or buffer. Dithiothreitol is available from Sigma Chemical Co., St. Louis, Cat # D0632. Dithioerythritol may be substituted.

 

DOPA (3-(3,4-Dihydroxyphenyl)-L-alanine MW 197.19)

8 mM	Dissolve 158 mg of L-DOPA to a final volume of 100 ml with water or buffer. Note that the maximum solubility of DOPA in water is 165 mg/100 ml (8.3 mM).

 

EDTA (Ethylenediaminetetraacetic acid MW 292.24)

1 M

Dissolve 292.24 grams of EDTA, free acid to a final volume of 1 liter. If the more soluble disodium salt of EDTA is used, adjust the weight accordingly. The pH can be adjusted with acetic acid or NaOH. For corresponding concentration dilutions, multiply the weight in grams by the desired molarity. For exam ple, for 10 mM EDTA, multiply 292.24 X 0.010 to obtain 2.92 grams of EDTA per liter.

 

EGTA (Ethylene Glycol-bis( aminoethyl Ether) N,N,N',N'-Tetraacetic Acid MW 380.4)

1 mM	issolve 380 mg of EGTA to a final volume of 1 liter with water or buffer.

 

Eosin

0.5% (w/v)

Dissolve 0.5 grams of Eosin Y in 100 ml of water.

 

Ethanol (C H OH MW 46.07)

Density = 0.7893 gm/ml

50-95% (v/v)

Since 95% ethyl alcohol is less expensive and easier to store than absolute, these dilutions should be made with 95% ethyl alcohol. Unless otherwise stated, denatured alcohol works as well as the more expensive non-denatured. A simple way to make the % so lution is to use the appropriate amount of 95% ethanol and dilute to 950 ml instead of 1 liter. For example, to make a 50% (v/v) solution, measure out 500 ml of 95% ethyl alcohol and dilute to a final volume of 950 ml with water. For a 70% solution, meas ure 700 ml of ethyl alcohol and dilute to 950 ml with water. Absolute ethanol should be used directly as 100% ethanol. It is important for histology that this be truly 100%. Since it is hydroscopic (it absorbs water from the air), do not assume it is abso lute unless it is sealed or treated to ensure no water. To test, add a drop to a sample of xylol. If any cloudiness occurs, the alcohol is not absolute.

8.5 M

The ethanol can be weighed (391.6 grams of absolute, 412.2 grams of 95% (v/v) or measured volumetrically by using the density. That is, 391.6 grams  0.7893 grams/ml or 496.1 ml of absolute ethanol. Using 95%, 412.2 grams  0.7893 grams /ml or 522.2 ml. Weigh or measure the appropriate amount and dilute to a final volume of 1 liter with water.

 

Ethanol-acetic acid fixative for histology (3:1)

To 75 ml of absolute alcohol, add 25 ml of glacial acetic acid. Must be made fresh, just prior to use.

 

Fetal Calf Serum (FCS)

While it is possible to prepare your own serum from whole blood, it is easier (and safer) to purchase FCS from a reputable supplier. Commercial sources are free of mycoplasma, pre-sterilized and controlled for the presence of antibodies. There are a n umber of serum substitutes available on the market and these may be less expensive when storage is considered. Suppliers include Gibco, Flow Laboratories, KC Biological and Sigma Chemical Co.  

 

Folin-Ciocalteu Reagent

This is usually purchased premixed, since it is difficult to make. Also known as 2N Folin-phenol reagent.

 

Giemsa stain

Prepare a stock solution by dissolving 3.8 grams of giemsa powder in 25 ml of glycerin. Heat gently with stirring for about 2 hours at 60 ° C. Cool and add 75 ml of methanol (neutral, acetone free).
For a working solution, dilute the stock solution 1/10 with water before use.
For chromosome banding, combine 5.0 ml of stock Giemsa, 3.0 ml of absolute methanol, 3.0 ml of 0.1 M citric acid and 89 ml of distilled water. Adjust the pH of the solution to 6.6 with Na HPO .

 

Glucose (MW 180.16)

10% (v/v)

Dissolve 10 grams of D-glucose (dextrose) in a final volume of 100 ml with water or buffer.

 

Glutaraldehyde (GTA)

5 %	GTA is usually supplied as a 25% or 50% (w/v) solution. It is used for electron microscope fixation as a 5% solution in a buffer. For routine use, add 20 ml of 25% GTA to 80 ml of 0.2 M sodium cacodylate buffer, pH 7.4.

 

Glycerol (MW 92.09)

10% (v/v)	To 10 ml of glycerol (glycerine) add enough water to make a final volume of 100 ml.
8 M	Weigh 73.67 grams of glycerol and add to a final volume of 100 ml. Alternatively, measure 499.1 ml of glycerol and make to a final volume of 1 liter (the density of glycerol at room temperature is 1.476) with water or buffer. For 8 M glycerol in MT buffer, make a 2X MT buffer for use as the diluent.