Heterologous probe a problem in Southern

spyong at pc.jaring.my spyong at pc.jaring.my
Sat Mar 23 06:26:37 EST 1996


Dear Netters,

I am trying to detect a particular gene from palm. I have a similar cloned
gene from maize as the probe. I realize these are two different species.
However, the sequence are pretty conserve. My Southern has failed for the
(to shameful to tell the no. of times!) time. 

I have a low 50C Hyb temp. and wash at the lowest of stringency..20X SSC.  I
use P32. My Hyb Buffer contains Dextran Sulphate, denatured Salmon
DNA,6XSSC, and blocking reagent know I transfred and the method is okay
because the only thing that lights up is the probe itself as a positive control.

Please can anybody shed some light? My project lies heavily on this detection.

P/S:  Now,I am detecting through PCR, but the bands that I get is a little
fuzzy. Is it okay to re-PCR the product?

Thanks a million!

My e-mail : spyong at pc.jaring.my

Stephanie.




More information about the Methods mailing list