Technique / Molecular Biology / DNA analysis techniques / DNA southern blot


Hybridization buffer recipe



Hybridization buffer recipe

Does anybody out there know a successful recipe of hybridization
buffer without the denatured salmon sperm DNA for genomic southern
hybridization?
Thanks.
XY Zheng
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Sure... use your usual hyb buffer (5X SSC, 10X Denhardts, 0.05% Na
pyrophosphate) and just boost your SDS to above 1%. For southerns I
normally ue 2% and haven't needed salmon sperm for over 6 years...


The old genescreen manual clued me onto this...
Hope this helps,
David
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This works fine for me.

7% SDS
0.5M Sodium Phosphate pH 7.2
1mM EDTA
1% BSA

I had no problem. Good luck.
N Kaushik
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Last update 26-Jul-2008, Rating of 2 votes.

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By Suresh on 19-Feb-2009
Hi, can you please send me a soft copy of complete protocol with reagents recipes and equipment for southern blot. I am new to the procedure. so, I will appreciate as many details as possible. My eid is drsureshvet@yahoo.co.in Thanks. Suresh
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By manju on 26-Jul-2008
i also had the same doubt. but i dono wat is the real reason to use Denatured solmon sperm DNA
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