User 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) per 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

(Bio-Rad Laboratories 1987) (Maniatis et al. 1982) (Thomas 1980)

1 mM EDTA 6x SSC 50% formamide

7% SDS 0.5% SDS 5x SSC

0.5 M NaHPO

, pH 7.2 5x Denhardt’s solution 1x Denhardt’s solution

100 µg/ml denatured 50 mM NaHPO

, pH 6.5

salmon sperm DNA 250 µg/ml denatured

1 mM EDTA salmon sperm DNA

For DNA or RNA Bound to For DNA Bound to For RNA Bound to

Zeta-Probe Membrane Nitrocellulose Nitrocellulose

A. Wash two 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 two times for 30–60 B. Wash at room B. Wash two times at

minutes at 65°C in: temperature for 50°C in:

1 mM EDTA, 5 minutes in: 2x SSC, O.5% SDS

0.1x SSC, 0.1% SDS, 1 mM EDTA

40 mM NaHPO

, pH 7.2,

1% SDS

DNA Bound to DNA Bound to

Zeta-Probe Membrane 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