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.
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