Solutions for molecular cloning--1 ) 2 ) 3 ) 4 ) 5 ) 6 ) 7 ) 8 ) 9 )
20X SSC (standard saline-citrate):
17.53 g NaCl
8.82 g sodium citrate
Dissolve in approx. 80 ml ddH2O, adjustpH to 7.0 with hydrochloric acid (HCl) and bring final volume to 100 ml.
1X SSC (standard saline-citrate):
5 ml 20X SSC
95 ml ddH2O
100 ml
5X Taq reaction buffer: 400 mM Tris-HCl, pH 9.0, 100 mM ammonium sulfate ((NH4)2SO4), pH 9.0, 25 mM MgCl2, and 5% dimethyl
sulfoxide (DMSO) in sterile double distilled water.
16
ml 1 M Tris-HCl,
pH 9.0
4 ml 1 M (NH4)2SO4, pH 9.0
1 ml 1 M MgCl2
2 ml DMSO
17 ml ddH2O
40 ml
5X Taq dilution buffer: 400 mM Tris-HCl, pH 9.0, 100 mM (NH4)2SO4, pH 9.0, and 25 mM MgCl2 in sterile double distilled water.
16 ml 1 M Tris-HCl, pH 9.0
4 ml 1 M (NH4)2SO4, pH 9.0
1 ml 1 M MgCl2
19 ml ddH2O
40 ml
5X Taq "A" termination mix: 62.5 uM dATP, 250 uM dCTP, 375 uM c7dGTP, 250 uM dTTP and 1.5 mM ddATP in 5 mM Tris-HCl, pH 7.6 and
0.1 mM EDTA.
20 ul 20 mM dATP
80 ul 20 mM dCTP
240 ul 10 mM 7deaza-dGTP
80 ul 20 mM dTTP
1920 ul 5 mM ddATP
640 ul 50:1 TE buffer
3420 ul sddH2O
6.4 ml
5X Taq "C" termination mix: 250 uM dATP, 62.5 uM dCTP, 375 uM c7dGTP, 250 uM dTTP and 0.75 mM ddATP in 5 mM Tris-HCl, pH 7.6
and 0.1 mM EDTA.
80 ul 20 mM dATP
20 ul 20 mM dCTP
240 ul 10 mM 7deaza-dGTP
80 ul 20 mM dTTP
960 ul 5 mM ddCTP
640 ul 50:1 TE buffer
4380 ul sddH2O
6.4 ml
5X Taq "G" termination mix: 250 uM dATP, 250 uM dCTP, 94 uM c7dGTP, 250 uM dTTP and 0.125 mM ddGTP in 5 mM Tris-HCl, pH 7.6
and 0.1 mM EDTA.
160 ul 20 mM dATP
160 ul 20 mM dCTP
120 ul 10 mM 7deaza-dGTP
160 ul 20 mM dTTP
320 ul 5 mM ddGTP
1280 ul 50:1 TE buffer
10600 ul sddH2O
12.8 ml
5X Taq "T" termination mix: 250 uM dATP, 250 uM dCTP, 375 uM c7dGTP, 62.5 uM dTTP and 1.25 mM ddTTP in 5 mM Tris-HCl, pH 7.6
and 0.1 mM EDTA.
160 ul 20 mM dATP
160 ul 20 mM dCTP
480 ul 10 mM 7deaza-dGTP
40 ul 20 mM dTTP
3200 ul 5 mM ddTTP
1280 ul 50:1 TE buffer
7480 ul sddH2O
12.8 ml
20X TAE buffer: 0.8 M Tris, 0.4 M NaOAc, and 0.04 M Na2EDTA, and glacial acetic acid to pH 8.3 in double distilled water.
96.9 g Tris base
32.8 g NaOAc-3H2O
14.9 g Na2EDTA
Dissolve in approx. 700 ml of double distilled
water, adjust the pH to 8.3 with glacial acetic acid, and bring to 1 L with ddH2O.
TEMED (N,N,N',N'-tetramethylethylenediamine): Kodak T-7024,
store protected from light at 15degC.
10xTB Salts:
2.31 g KH2PO4
12.54 g K2HPO4 (potassium phosphate, dibasic)
ddH2O to
100 ml autoclave)
Terrific Broth (TB):
12 g Bacto-tryptone
24 g yeast extract
4 ml glycerol
ddH2O to 900 ml
Autoclave, cool and add 100 ml of 10xTB Salts and adjust the final volume to 1 L with sddH2O.
TE (10:0.1) buffer:10
mM Tris-HCl, pH 7.6, 0.1 mM EDTA
10 ml 1 M Tris-HCl, pH 7.6
0.2 ml 0.5 M EDTA
ddH2O to 1 L
TE (10:1) buffer: 10 mM Tris-HCl, pH 7.6, 1 mM EDTA
10 ml 1 M Tris-HCl, pH 7.6
2 ml 0.5 M EDTA
ddH2O to 1 L
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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