Manual

50% Formamide

Dilute 50.0 g formamide to 100 ml with ddH

O. 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

, 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

, 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