Bio-Rad Bio-Dot® and Bio-Dot SF Microfiltration Apparatus - Manual (Page 21)

Section 9
Solutions for Nucleic Acid Applications
For DNA or RNA Bound to For DNA Bound to For RNA Bound to
Zeta-Probe Membrane Nitrocellulose Nitrocellulose
1 mM EDTA 6x SSC 50% formamide
7% SDS 0.5% SDS 5x SSC
0.5 M NaHPO
4
, pH 7.2 5x Denhardt’s solution 1x Denhardt’s solution
100 µg/ml denatured 50 mM NaHPO
4
, pH 6.5
salmon sperm DNA 250 µg/ml denatured
1 mM EDTA salmon sperm DNA
20x SSC
3 M NaCl
0.3 M trisodium citrate (FW = 294.1)
Dissolve 175.0 g NaCl and 88.2 g trisodium citrate in ddH2O.
Adjust volume to 1 L with ddH2O.
20x SSPE
3.6 M NaCl
0.2 M Na2HPO4 •7H2O (FW = 268.07)
0.02 M EDTA
Dissolve 210.0 g NaCl, 53.6 g Na2HPO4 •7H2O, 7.44 g EDTA in ddH2O.
Adjust volume to 1 L with ddH2O.
TE
10 mM Tris-HCl, pH 8.0
1 mM EDTA, pH 8.0
Dilute 10 ml 1 M Tris-HCl, pH 8.0 and 4 ml 0.250 M EDTA, pH 8.0 to 1 L with ddH2O.
100x Denhardt’s Solution
2% bovine serum albumin (BSA)
2% polyvinylpyrrolidone
2% Ficoll
Dissolve 2.0 g BSA, 2.0 g polyvinylpyrrolidone, 2.0 g Ficoll in ddH2O.
Adjust volume to 100 ml with ddH2O.
20% SDS
Dissolve 20.0 g SDS in ddH2O. Adjust volume to 100 ml with ddH2O.
It may be necessary to heat to 65°C to get into solution.
1 M NaHPO4, pH 7.2
1 M Na2HPO4 •7H2O (FW = 268.07)
Dissolve 134.0 g Na2HPO4 •7H2O in dd H2O. Add 4 ml 85% H3PO4.
Adjust volume to 1 L with ddH2O.
50% Dextran Sulfate
50% dextran sulfate
0.2% sodium azide
Dissolve 50.0 g dextran sulfate and 0.2 g sodium azide in ddH2O.
Adjust volume to 100 ml with ddH2O. Store at 4°C.
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