Reagent and Buffer Recipes

20x SB Agarose Gel Buffer (Sodium Borate)

38.2 g Borax
10 g Boric Acid
to 1 L ddH20

Mix with a stir bar until everything is dissolved and liquid is clear. Pour into one of the 20 L carboys and fill to 20 L with ddH20 to make it 1x.

6x Bromophenol Blue Gel Loading Buffer

30 ml Glycerol
0.01 g Bromophenol Blue
to 50 ml ddH20

DNA Ladder with Loading Dye

375 ul Loading Buffer (above)
250 ul Ladder
325 ul 10x PCR Buffer
3 ml ddH20

Makes ~4ml. Aliquot to 1.5 ml centrifuge tubes.

Stock solutions

Tris-HCl, 1 M

121 g Tris base in 800 ml ddH20
Adjust to pH 8.0 with HCl
Mix and add ddH20 to 1 L

EDTA, 0.5 M

Dissolve 186.1 g NaEDTA 2 dH20 in 700 ml ddH20
Adjust pH to 8.0 with 10 M NaOH (~ 50ml)
Mix and add ddH20 to 1 L

NaOH, 10 M

Dissolve 400g NaOH in 450 ml ddH20
Mix and add ddH20 to 1 L

Potassium acetate, 5 M

29.5 ml glacial acetic acid
KOH pellets to pH 4.8 (several)
ddH20 to 100 ml
Store at room temperature

NaCl, 1M

Dissolve 58.4 g of NaCl in 800 ml ddH20
Mix and add ddH20 to 1 L

CaCl, 1 M

Dissolve 110.9 g of CaCl in 800 ml ddH20
Mix and add ddH20 to 1 L
Aliquot into 2 500 ml bottles and autoclave

MgSO4, 1 M

Dissolve 120.3 g of MgSO4 in 800 ml ddH20
Mix and add ddH20 to 1 L
Aliquot into 2 500 ml bottles and autoclave

RNase A, 5 mg/ml

Dissolve 100 mg of RNase A in 20 ml of 0.05% glacial acetic acid, and transfer to a 50-ml conical tube
Place the tube in a boiling-water bath for 15 minutes
Cool the solution, and neutralize by adding 120 μl of 1 M Tris (pH 8.0)
Distribute 1 ml aliquote into 1.5 ml MFT, and store at -20 C
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Topic revision: r5 - 09 Sep 2011 - 15:51:54 - CraigBarnhart
 
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