To make 4 ml of these nucleotides at a final concentration of 0.25 mM from the stock 100 mM solutions, add the following:

10 ul 100 mM dATP

10 ul 100 mM dCTP

10 ul 100 mM dGTP

10 ul 100 mM dTTP

3.6 ml ddH₂O

Aliquot into 0.5 eppendorf tubes with 10 ul in each tube.

To order these nucleotides, call Pharmacia at 1 800-526-3593. Order the dNTP set: 27-2035-01 dNTP set (100mM each dATP, dCTP, dGTP, dTTP- each in 250 ul volume) $174.00 for the set.

20% glucose:

20 g d-glucose

ddH₂O to 100 ml

filter sterilize

6 M guanidine HCl, pH 6.4, containing 50 mM Tris-HCl, 20 mM EDTA:

573.18 g guanidine-HCl (Sigma G-4505)

50 ml 1 M Tris-HCl, pH 7.6

40 ml 0.5 M EDTA, pH 8.0

ddH₂O to 1 liter

GET/lysozyme solution: (50 mM glucose, 25 mM Tris-HCl, pH 8.0, and 10 mM EDTA, pH 8.0 in double distilled water)

0.9 g d-glucose

2.5 ml 1 M Tris-HCl, pH 8.0

2 ml 0.5 M EDTA, pH 8.0

ddH₂O to 100 ml (filter sterilize and store at 4degC).

Add 2 mg/ml lysozyme (Sigma L-6876) just before use.

1 M HEPES, pH 7.5:

23.83 g HEPES (Sigma H-3375)

ddH₂O to 100 ml

adjust pH to 7.5 with potassium hydroxide (KOH) (store at 4deg.C).

上一页  [1] [2] [3] [4] [5] [6] [7] [8] [9] [10]  ... 下一页  >>