How do you manage your working sample DNA in RAPD?

Klaus.Matthaei at ANU.EDU.AU Klaus.Matthaei at ANU.EDU.AU
Wed Jun 1 00:06:10 EST 1994


>Hello netters,
>
>I am having difficulty in the Abalone DNA. Once the DNA put into -20 or -70
>or 4C frige, there was NO RAPD bands immediately. I presume that the DNA
>was broken down during the frozen processing. Normally how do you arrange
>the DNA if you will use the same DNA in several RAPD reactions?
>
>Heart-felt thanks.
>
>With regards.
>
>Bixing Huang
>huangbx at deakin.edu.au

This is NOT a flame

Why is it that so many people freeze everything at -20??

I have kept genomic DNA (mammalian) at 4*C for YEARS (ALMOST DECADES) in
T10 E1 i.e Tris 10 mM EDTA 1mM (more recently 0.1mM EDTA) without ANY
problems of amplifying single copy genes.

WHY FREEZE THAW??

I find that freeze-thawing degrades (sheares) the DNA and can give very
poor PCR results while at the same time does not appreciably do anything to
Southern analysis except weaken the signal.

Please: store at 4*C

The DNA survives much better.

my $0.02 worth

Cheers, Klaus

*************************************************************************
Dr Klaus Matthaei                               |           |
The John Curtin School of Medical Research      |  _--_|\   |
The Australian National University              | /      \  |
Snail mail: Canberra, ACT 0200, Australia       | \_.--._/<<|
E-mail: Klaus.Matthaei at anu.edu.au               |       v   |
Landline: 61 6 249 3782
                        "Sometimes I'm the Louisville slugger"
                        "Sometimes I'm the Ball"
                                Dire Straits
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