How to make stock solutions? BSA, EtBr, 0.5 M EDTA, 0.5 M Na3EDTA, Polyethylene Glycol, Perchloric acid, 1M NaHPO4, Tris 1 M, Treacking Dye, Glycerol Solution, RNase A and RNase T1

Stock Solution

BSA

Stock BSA from BRL comes as 50 mg/1000 �l. Add 4 ml of H2O for a final concentration of 10 mg/ml. Store at -20 C.

EtBr (10 mg/ml)

Dissolve 100 mg EtBr in 10 ml H2O. Stir well and store at 4 C in an amber bottle.

0.5 M EDTA (pH 8.0): 500 ml

Dissolve 95.05 g of Na2EDTA in appr. 400 ml H2O. Adjust pH to 8.0 with 6 N NaOH. Then bring volume to 500 ml and autoclave.

0.5 M Na3EDTA: 1L

Add 186.2 g (?) to 900 ml H2O. Adjust pH with conc HCl (or NaOH ?) to 8.0, and bring up to 1 L.

5 M LiCl 50 mM MOPS (pH 8.0)

Conc
5 M LiCl 21.2 g
50 mM MOPS 1.05 g
H2O up to 100 ml
Adjust pH 8.0 with NaOH. Store at 4 C.

50 % PEG

Add 100 g PEG (Polyethylene Glycol, appr. MW 8.000) to 50 ml H2O and stir for about one hour. Then bring up to 200 ml with H2O.

3 N Perchloric acid

Dilute 49.2 ml of perchloric acid up to 200 ml with H2O in a hood. Stable at room temperature for several month. Treat with care.

1 N NaOH

Dissove 40 g of NaOH in 1000 ml H2O.

1M NaHPO4

Stock
NaH2PO4.H2O (monobasic monohydrate) 47.5 g
Na2PO4.7H2O (dibasic 7-hydrate) 42.2 g
H2O to 500 ml

10 M NH4Ac

Add 400 ml H2O to a 1 Kg bottle NH4Ac, dissolve, and adjust to 1300 ml.

Tris 1 M pH 8.0

Stock
Tris-HCl 88.8 g 177 g 355.2 g
Tris-Base 53.0 g 106.0 g 212.0 g
H2O 1L 2 L 4 L

Tris 1 M pH 7.5

Stock
Tris-HCl 127 g 254 g 508 g
Tris-Base 23.6 g 47.2 g 94.4 g
H2O 1 L 2 L 4 L

76 % EtOH 10 mM NH4Ac

Conc. Stock Volume
75 % 95 % EtOH 80.0 ml
0.2 M 5 M NH4OAc 0.2 ml
H2O 19.8 ml
5 M NH4Ac stock = 38.55 g/100 ml

76 % EtOH 0.2 M NaAc

Conc. Stock Volume
76 % 95 % EtOH 160 ml
0.2 M 3 M NaAc 13.33
H2O 26.67
3 M NaOAc stock = 40.83 g/100 ml

0.2 N NaOH 0.6 M NaCl

1L 2L 3L 4L
NaOH 8.0 g 16.0 g 24.0 g 32.0 g
NaCl 35.04 g 70.08 g 105.2 g 140.2 g

0.5 M Tris (pH 7.5) 1.5 M NaCl

1L 2L 3L
Tris-HCl 63.5 g 127.0 g 190.0 g
Tris-Base 11.8 g 23.6 g 35.4 g
NaCl 87.6 g 175.0 g 262.8 g

25 mM NaHPO4

1M NaHPO4 100 ml
H2O to 4 L

10 X Treacking Dye

Conc.
50 mM Tris pH 8.0
50 mM Na3EDTA
25 % Glycerol
5 % Ficoll
0.1 % Xylenol Cyanol
0.1 % Bromophenol Blue
1 % SDS

Glycerol Solution

Glycerol 650 ml
1 M MgSO4 100 ml
1 M Tris (pH 8) 25 ml
H2O up to 1 L
Mix well and autoclave for 15 min at 121 C.

0.1 M Spermidine Solution

Dissolve 255 mg spermidine (Sigma #S2501) in H2O to a final volume of 10 ml. Store at -20 C.

Saline-MOPS

1X 1L 1X
0.85 % NaCl 8.5 g
50 mM MOPS 6.3 g MOPS free acid
4.7 g MOPS Na salt
pH 7 autoclave and store at room temperature.

Proteinase K - 10 mg/ml in TE

Weigh out 10 mg proteinase K (Boeringer Manheim #161 497) and dissolve in 1 ml TE. The solution can be used immediately or aliquoted and stored at -20 C

RNase A and RNase T1

Dissolve 100 mg RNase A (bovine pancrease: Sigma #R4875), if desired. add 5,000 units of RNase T1 together, in 10 ml of 10 mM Tris 15 mM NaCl. Boil for 15 min and allow to cool slowly to room temperature. Distribute 1 ml aliquote into 1.5 ml MFT, and store at -20 C. If