Help! my colony blots are lighting up like a christmas tree

R. Jayakumar jakku at mrna.tn.nic.in
Wed Feb 16 10:27:48 EST 2000


Hi
   Somebody should really help me out with this problem.  I did colony lifts
of about 1000 clones onto Both nitrocellulose and Nylon (N+) colony lift
membrane discs and did a normal hybridisation with my DNA probe labelled
with p32 using the GIBCO BRL's random primer labelling kit.  But nearly all
my colonies lighted up, with both the membranes I used.
    Can anybody tell me where the mistake is?  For your information, I
followed the colony lift technique given in Maniatis manual.  As
recommended, I have also given prewashes of the blot before doing
hybridization.
    Any suggestion will be most welcome
thanking you in advance

cheers
jayakumar

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R. JAYAKUMAR
CSIR- Senior Research Fellow
Dept. of Molecular Microbiology,
School of Biotechnology,
Madurai Kamaraj University,
Madurai - 625 021.
India
email: jakku at usaf.org
tel: +91-452-858471-374
efax: (603)-688-4665
web: http://members.tripod.com/~jakspage
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"There is a possible even in imPOSSIBLE"


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