Gene copy number by slot-blot hybridization

R. Jayakumar jakku at mrna.tn.nic.in
Fri Jan 28 13:54:56 EST 2000


just try placing the nylong membrane after transfer in  a microwave oven at
full setting (atleast 800W) for 2.45 min.  This came in a NAR paper around
1991.  I don't remember the exact reference, but it gives excellent fixing
and much better signals than UV crosslinking or hot oven fixing at 80C,
        try this out.
bye
cheers
jakku
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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"

----- Original Message -----
From: John Lum <johshi at hotmail.com>
To: <methods at hgmp.mrc.ac.uk>
Sent: Friday, January 28, 2000 10:08 AM
Subject: Re: Gene copy number by slot-blot hybridization


> Hi,
>
> >     How do you fix the DNA on the membrane? Sometimes, just drying the
membrane
> > would be enough; but if you want a better fixation you may try to use UV
> > irradiation or heat fixation (placing the membrane in an incubator at
80ºC for
> > 1.5-2 hours would do both a partial denaturation of the DNA and fixation
to the
> > membrane).
>
>       I've try both UV-fixation and baking at 120C for 40 mins. Both of
them have
> similar result.
>
> Regards,
> Lum
>
>

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