Bio-Rad Bio-Dot® and Bio-Dot SF Microfiltration Apparatus - Manual (Page 22)

50% Formamide
Dilute 50.0 g formamide to 100 ml with ddH2O. Store at 4°C. Immediately before use, deionize the
required volume by stirring gently for 1 hour with 1 g mixed bed ion exchange resin (AG ® 501-X8 (D)
resin, catalog number 142-6425)/10 ml of formamide. Filter through coarse filter paper.
For DNA or RNA Bound to For DNA Bound to For RNA Bound to
Zeta-Probe Membrane Nitrocellulose Nitrocellulose
A. Wash 2 times for 30–60 A. Rinse in: A. Wash 4 times at room
minutes at 65°C in: 2x SSC, 0.5% SDS temperature for 5 minutes
1 mM EDTA in 2x SSC, 0.1% SDS
40 mM NaHPO
4
, pH 7.2
5% SDS
B. Wash 2 times for 30–60 B. Wash at room B. Wash 2 times at 50°C in:
minutes at 65°C in: temperature for 5 minutes 0.1x SSC, 0.1% SDS
1 mM EDTA in: 2x SSC, 0.5% SDS
40 mM NaHPO
4
, pH 7.2
1% SDS
C. Wash at 68°C 2 times
for 1 hour each in:
0.1x SSC, 0.5% SDS
DNA Bound to Zeta-Probe Membrane DNA Bound to Nitrocellulose
50% formamide 50% formamide
1.5x SSPE 1x Denhardt’s solution
1% SDS 0.1% SDS
0.5% nonfat dry milk 100 µg/ml denatured salmon sperm DNA
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