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RNA/Zeta Probe Dot Blotting protocol

RNA/Zeta Probe Dot Blotting protocol

1) Make up RNA (up to 20ug) dissolved in sterile H2O, TE or 0.5% SDS to 500ul with ice-cold sterile 10mM NaOH, 1mM EDTA and apply it to Zeta Probe membrane, held in a dot-blot apparatus, which has been pre-wetted in 2 x SSC.

2) Apply a vacuum across the apparatus to draw the solution through the membrane and wash each dot once with 500ul ice-cold 10mM NaOH, 1mM EDTA.

3) Remove membrane from apparatus and wash for 5 mins in 2 x SSC at room temperature.

4) Allow membrane to air dry.

At this stage the membrane can be stored dry at -20°C until required for hybridisation to the probe of choice or it can be used immediately.

Keywords: RNA, dot blot
Contact Email Address: Simon.Dawson@nott.ac.uk
Submitted at 16:58 on 22/1/96 by:

Dr. Simon Dawson,
Department of Biochemistry,
University of Nottingham,
The Medical School ,
Q.M.C. ,
Clifton Boulevard ,
Nottingham,
NG7 2UH,
U.K..
Tel: +44 (0)115 9249924 ex.44787,
FAX: +44 (0)115 9422225,

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