SSC conc. in prehyb.
ckwen at expert.cc.purdue.edu
Fri Feb 21 14:44:26 EST 1997
Denni Schnapp (ds4 at st-andrews.ac.uk) wrote:
: Hi folks,
: We prehybe our filters (in my case it was Hybond-C nitrocellulose) in 6 x SSC,
: 0.5% SDS, 5X Denhards's (I think that is right) final conc., plus salmon sperm
: DNA. Like a moron, I copied the corresponding recipe down wrongly and ended up
: with 5 x SSC. Now, some prehyb.-buffers appear to contain 5 x SSC, others contain 6 x SSC. What are peoples experience with the SSC conc.?
: Thanks for any input,
: Denni (ds4 at st-and.ac.uk
I would not be worried. 5X SSC contains about 0.825M of salt, while 6XSSC
contains about 0.99M of salt. I always use Church buffer which contains 0.5M
of salts. For Church buffer, I always get clear results, which means that under
5XSSC conditions, you would not have problems in hybridization.
Furthermore, the decrease in Tm can be estimated by
caculating the differences between
log[6xssc] and log[5xssc], and I don't think that be significant.
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