Parameters for Southern Hybs!
sweechie at HOTMAIL.COM
Mon Jun 14 21:06:21 EST 1999
I am having a lot of trouble with southern hybs of genomic DNA from CHO
cells.The fragments I am trying to resolve are 2000,2500,5000,7500bp
long.The 2500bp and 5000bp fragments resolve however not the higher
molecular weight fragment.In my gel washing step,I depurinate the gel by
immersing it for 20 minutes or so,in a solution of 0.25 HCl. would that be
too long or too short a time and could that be the reason why my bands are
coming up? Also,I am having to expose the film for at least 10-12 day before
any of the CHo clones come up.Could the time of exposure be shortened in any
way, say if I didn't add formamide to my hyb mix or if i used intensifying
screens.Any suggestions would be great.
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