Solutions for molecular cloning--1 ) 2 ) 3 ) 4 ) 5 ) 6 ) 7 ) 8 ) 9 )
1 M MOPS:
20.93 g MOPS (Sigma M-1254)
Dissolve in 80 ml ddH2O, adjust pH to 7.5 with 1 N NaOH, and bring volume to
100 ml.
10X MOPS buffer: 400 mM MOPS, pH 7.5, 500
mM NaCl, 100 mM MgCl2 in double distilled water.
400 ul 1 M MOPS, pH 7.5
170 ul 3 M
NaCl
100 ul 1 M MgCl2
330 ul ddH2O
1 ml
2.7 M MOPS (acid form):
5.65 g MOPS (acid form)
ddH2O to 10 ml
MOPS-Acid buffer: 1.35 M MOPS (acid form), 100 mM MgCl2 in double distilled water.
500 ul 2.7 M MOPS (acid form)
100 ul 1 M MgCl2
400 ul ddH2O
1 ml
10X Mn2+/isocitrate buffer: 50 mM MnCl2, 150 mM isocitrate (Na salt), 25% glycerol in double distilled water
50 ul 1 M MnCl2
150 ul 1 M isocitrate
250 ul glycerol
550 ul ddH2O
1 ml
10x MTBE (Modified Tris-borate-EDTA buffer): 1.3 M Tris, 0.4 M Boric acid, and 0.02 M EDTA in double distilled water.
162 g Tris base
27.5 g Boric acid
9.3 g Na2EDTA
ddH2O to 1 L
Nucleotide ordering information:
100 mM dATP 27-2050-01 Pharmacia
100 mM dCTP 27-2060-01 Pharmacia
100 mM dGTP 27-2070-01 Pharmacia
10 mM c7dGTP 988 537 Boehringer-Mannheim
100 mM dTTP 27-2080-01 Pharmacia
5 mM ddATP 27-2057-00 Pharmacia
5 mM ddCTP 27-2065-00 Pharmacia
5 mM ddGTP 27-2075-00 Pharmacia
5 mM ddTTP 27-2085-00 Pharmacia
20 mM dNTP stocks: Prepare from 100 mM stocks
80 ul 100 mM dNTP
40 ul 50:1 TE buffer
280 ul ddH2O
400 ul
5 mM dNTP stocks: Prepare from 20 mM stocks
25 ul 20 mM dNTP
10 ul 50:1 TE buffer
65 ul ddH2O
100 ul
2 mM dNTPs: 2 mM dATP, dCTP, dGTP, and dTTP in 5 mM Tris-HCl, pH 7.6, 0.1 mM EDTA
100 ul 20 mM dATP
100 ul 20 mM dCTP
100 ul 20 mM dGTP
100 ul 20 mM dTTP
100 ul 50:1 TE buffer
500 ul ddH2O
1 ml
2 mM [alpha]-S-dNTPs: 2 mM [alpha]-S-dATP, [alpha]-S-dCTP, [alpha]-S-dGTP, and [alpha]-S-dTTP in 5 mM Tris-HCl, pH 7.6, 0.1 mM
EDTA
100 ul 20 mM [alpha]-S-dATP
100 ul 20 mM [alpha]-S-dCTP
100 ul 20 mM [alpha]-S-dGTP
100 ul 20 mM [alpha]-S-dTTP
100 ul 50:1 TE buffer
500 ul ddH2O
1 ml
3M NaCl (sodium chloride):
17.53 g NaCl
ddH2O to 100 ml
10N NaOH (sodium hydroxide):
40 g NaOH
ddH2O to 100 ml.
1N NaOH:
10 ml 10 N NaOH
ddH2O to 100 ml
9.5M NH4OAc (ammonium acetate):
73.23 g NH4OAc
ddH2O to 100 ml
8.0M NH4OAc:
61.69 g NH4OAc
ddH2O to 100 ml
10X PCR buffer: 500 mM KCl, 100 mM Tris-HCl, pH 8.5, 15 mM MgCl2 in sterile double distilled water
5 ml 1 M KCl
1 ml 1 M Tris-HCl, pH 8.5
150 ul 1 M MgCl2
ddH2O to 10 ml
PCR Deoxynucleotide Preparation: To make 12.5 ml of the PCR nucleotides at a concentration of 2 mM each nucleotide,
combine the following:
250 ul 100 mM dATP
250
ul 100 mM dCTP
250 ul 100 mM dGTP
250 ul 100 mM
dTTP
11.5 ml ddH2O
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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