lose in primer viability

Christine Schneider Christine.Schneider at ruhr-uni-bochum.de
Thu Apr 5 02:04:11 EST 2001


I think the EDTA might cause a problem in the PCR reaction. EDTA chelates (?) Mg,
which is necessary for DNA-Polymerase.

Christine


"Dr. Hiranya S. Roychowdhury" schrieb:

> Always reconstitute in a buffer. The concentration of the buffer may be as
> loww as 1mM, but the NA's need to be buffered for longterm storage. 10mM
> Tris.Cl, pH7.5 to pH8.0 is used almost universally. The TE may also be used.
>
> At 11:15 AM 4/4/01 +0100, R. Jayakumar wrote:
> >Hi.
> >I have a strange problem with one of my set of primers.  This pair of
> >primers is meant to amplify an antibiotic resistance gene.  the primers work
> >well only for two months or so, even though I use only a working stock and I
> >maintain the stock in -20C.  The primers somehow lose their viability after
> >2-3 months and I have to synthesize them again.  The same problem happens
> >again after 2-3 months.  I reconstitute the lyophilized primers in deionised
> >water as soon as I receive it and store in -20C.  This is becoming a
> >problem, because I have to synthesize them every so often.
> >    can anybody suggest what could be the problem?
> >sincerely
> >jayakumar
> >
> >||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
> >||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
> >||||
> >R. JAYAKUMAR
> >CSIR- Senior Research Fellow
> >Dept. of Molecular Microbiology,
> >School of Biotechnology,
> >Madurai Kamaraj University,
> >Madurai - 625 021.
> >India
> >email: jakku at linuxfan.com
> >tel: +91-452-858471-374
> >office fax(india)= +91-452-859105
> >Efax(USA): (603)-688-4665
> >||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
> >||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
> >||||
> >"Life is a comedy for those who think and a tragedy for those who
> >feel" --Emerson
> >
> >---
> >
> >
> Dr. Hiranya Sankar Roychowdhury
> College Asst. Prof.
> Molecular Biology,
> Dept. of Chemistry & Biochemistry
> Box 30001 - 3MLS
> New Mexico State University
> Las Cruces, NM 88003
>
> Lab: (505) 646 4722
> Office: (505) 646 8256
> hroychow at nmsu.edu
>
> ---

--
______________________________________________________________

Christine Schneider

Ruhr-Universität Bochum
AG Molekulare Genetik
ND 06/756
44780 Bochum

Tel.: 0234 32-25629
email: christine.schneider at ruhr-uni-bochum.de





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