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Re: southern hybridisation/blotting
Posted by: Jim (IP Hidden, Unregistered user, )
Date: September 14, 2005 10:16AM
Hi
I have been doing Southern hybridization for the last 6 years and routinely use 68C for hybridization and washing of the filters, when my probe is above 50 bp. I routinely use 1.5XSSC and 0.1%SDS for my washes, and get good clean signals. Very rarely, I have to use more stringent washes. For oligos I use 42C for hybridization and washing.
So in my mind 68C is ok for Southern in most cases.
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Re: southern hybridisation/blotting
Posted by: Jim (IP Hidden, Unregistered user, )
Date: September 14, 2005 10:18AM
Skipping this step will make your probe more likely to rehybridize to itself. Do not skip the step "place on ice" or you will most likely lose sensitivity.
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