Alkaline downward trnasfering

Gen Yan Yang gyang at MED.UNC.EDU
Sat Mar 6 21:58:33 EST 1999


Dear all, 
I use alkaline downward transfering method to transfer DNA to nylon N+
memberane. After depurinization in 0.25 N HCl for 15 min, I just rinse
the gel with D.W. instead of denaturation in o.4 N NaOH , then the gel
was transfered with 0.4 N NaOH for several hours. I do not know if the
DNAs in my gel have been changed into single strand during transfering.
is it necessary to denature the DNA before transfering since the
transfer buffer is 0.4 N NaOH?  Thanks!

Yang
the University of Northe Carolina at Chapel Hill.



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