Solutions for molecular cloning--1 ) 2 ) 3 ) 4 ) 5 ) 6 ) 7 ) 8 ) 9 )
10X ABI TBE:
216 g Tris base
110 g boric acid
16.6 g EDTA
Add water to 2 liters.
40% Acrylamide/Bisacrylamide (40%
A&B):
380 g Acrylamide (Kodak 5521)
20
g N,N-Methylene-bisacrylamide (Kodak 8383)
Dissolve in approx. 800 ml of doubledistilled water and then deionize by stirring with 50 g of Amberlite MB-1 (
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)
ddH2O 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)
ddH2O to 100 ml (make fresh)
15% Ammonium persulfate (APS):
1.5 g APS (Kodak 11151)
ddH2O to 10 ml (store at 4deg.C).
Ampicillin (Amp): Stock of 5 mg/ml in sterile ddwater (sddH2O).
0.5 g Amp (Sigma A-9518)
sddH2O to 100 ml (Add to media for final conc. 100 ug/ml)
100 mM rATP (adenosine triphosphate):
619 mg dipotassium ATP (ICN 100004)
sddH2O
to 10 ml (aliquot and store at -20deg.C).
1 mg/ml BSA (bovine serum albumin):
5 mg BSA (Sigma A-9647)
sddH2O to 5 ml (aliquot and store at -20deg.C)
Bst dilution buffer: 50 mM HEPES, pH 7.6, 10 mM MgCl2, 1 mM DTT, and 1 mg/ml
BSA in sterile double distilled water.
500 ul 1 M HEPES, pH 7.6
100 ul 1 M MgCl2
10 ul 1 M DTT
10 mg BSA
sddH2O to 10 ml
Bst reaction buffer: 500 mM Tris-HCl, pH 8.5 and 150 mM MgCl2 in sterile double distilled water.
5 ml 1 M Tris-HCl, pH 8.5
1.5 ml 1 M MgCl2
sddH2O to 10 ml
Bst nucleotide extension mix: 15 uM dCTP, 7deaza-dGTP, and dTTP in 5 mM Tris-HCl, pH 7.6 and 0.1 mM EDTA
3 ul 5 mM dCTP
3 ul 5 mM 7deaza-dGTP
3 ul 5 mM dTTP
100 ul 50:1 TE buffer
891 ul sddH2O
1 ml
Bst "short" termination "A" mix: 8 uM dATP, 164 uM dCTP, 164 uM 7deaza-dGTP, 164 uM dTTP, 660 uM ddATP in 5 mM Tris-HCl, pH 7.6
and 0.1 mM EDTA.
8 ul 0.5 mM dATP
16.4 ul 5 mM dCTP
16.4 ul 5 mM 7deaza-dGTP
16.4 ul 5 mM dTTP
66 ul 5 mM ddATP
50 ul 50:1 TE buffer
326.8 ul sddH2O
500 ul
aliquot (18 ul for 6 reactions) and store at -70degC
Bst "short"
termination "C" mix: 8 uM dCTP, 164 uM dATP, 164 uM 7deaza-dGTP, 164 uM dTTP, 400 uM ddCTP in 5 mM Tris-HCl, pH 7.6 and 0.1 mM EDTA.
16.4 ul 5 mM dATP
8 ul 0.5 mM dCTP
16.4 ul 5 mM 7deaza-dGTP
16.4 ul 5 mM dTTP
40 ul 5 mM ddCTP
50 ul 50:1 TE buffer
352.8 ul sddH2O
500 ul
aliquot (18 ul for 6 reactions) and store at -70degC
Bst "short"
termination "G" mix: 8 uM dGTP, 164 uM dATP, 164 uM dCTP, 164 uM dTTP, 540 uM ddGTP in 5 mM Tris-HCl, pH 7.6 and 0.1 mM EDTA.
16.4 ul 5 mM dATP
16.4 ul 5 mM dCTP
8 ul 0.5 mM 7deaza-dGTP
16.4 ul 5 mM dTTP
54 ul 5 mM ddGTP
50 ul 50:1 TE buffer
338.8 ul sddH2O
500 ul
aliquot (18 ul for 6 reactions) and store at -70degC
Commonly used solutions (1)--- 1 ) 2 ) 3 ) 4 ) 5 ) 6 )
Commonly used solutions (2)--- 1 ) 2 ) 3 )
Solutions for molecular
cloning---1 ) 2 ) 3 ) 4 ) 5 ) 6 ) 7 ) 8 ) 9 )
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