Thermo Scientific Shake 'n Stack 6244, Руководство по эксплуатации и техническому обслуживанию

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© Thermo
Scientific
, May 2003. Issue 7 24
4. Stringency washing is carried out as follows using large volumes (at least 50ml) of
the following solutions, pre-warmed to the required temperature:
2 x 15 minutes with 6 x SSPE
(SSC)
0.1% SDS at the Hybridisation
temp.
1 x 2 minutes with 6 x SSPE
(SSC) 0.1% SDS at the Tm
5. Wrap the membrane in Saran Wrap

and autoradiograph at -70
°
C in a cassette
with an intensifying screen. Expose initially for approximately 12 hours (or
overnight).
Background Hybridisation problems are much more common when using oligonucleotide
probes. For this reason procedures to reduce background such as pre-washing of
colony filters (Chapter 2, Colony/Plaque Blotting) and purification of the oligonucleotide
to remove unincorporated nucleotides are particularly advantageous.