Table of Contents

10X TBE

40% Acrylamide Stock

Alkaline lysis solution

10X TBE:

216 g Tris base

110 g boric acid

16.6 g EDTA

Add water to 2 liters.

40% Acrylamide/Bisacrylamide (40% A&B):

38 g Acrylamide (Kodak 5521)

2 g N,N-Methylene-bisacrylamide (Kodak 8383)

Dissolve in approx. 80 ml of double distilled water and then deionize by stirring with 5 g of Amberlite MB-1 (Sigma MB-1A) for 1 hour at room temperature. Suction filter to remove the Amberlite and adjust to a final volume of 100 ml with double distilled water. (store at 4deg.C).

10x Agarose gel loading dye:

15% Ficoll, 0.2% bromophenol blue, 0.2% xylene cyanol FF in double distilled water.

1.5 g Ficoll (Sigma F-2637)

0.02 g Bromophenol blue (Sigma B-0126)

0.02 g xylene cyanole FF (Kodak T-1579)

ddH₂ O to 10 ml (store at -20deg.C).

Alkaline lysis solution (NaOH/SDS) :

0.2 N NaOH, 1% SDS in ddwater.

20 ml of 1 N NaOH (or 0.8 gms)

10 ml of 10% SDS (or 1.0 gms)

ddH₂ O to 100 ml (make fresh)

15% Ammonium persulfate (APS):

1.5 g APS (Kodak 11151)

ddH₂ O to 10 ml (store at 4deg.C).

100 mM rATP (adenosine triphosphate) :

619 mg dipotassium ATP (ICN 100004)

sddH₂ O to 10 ml (aliquot and store at -20℃).

1 mg/ml BSA (bovine serum albumin) :

5 mg BSA (Sigma A-9647)

sddH₂ O to 5 ml (aliquot and store at -20℃)

100 mM calcium chloride (CaCl₂) :

1.48 g CaCl₂-2H₂ O

ddH₂ O to 100 ml

autoclave to sterilize (store at 4deg.C).