Marker

Arnoud van Vliet avvliet at knoware.nl
Thu Feb 24 16:46:14 EST 2000


Hi

your second band is probaly false priming. When I used the standard
eubacterial 16S primers for amplification, I also got two products. The
higher one sounds like the one to use. Just clone both of them, just to make
sure

good luck
Arnoud

"Ma Sheila M. De Jesus" <mshmdeje at USTCC.ust.edu.ph> wrote in message
news:Pine.OSF.3.96.1000224115918.5740B-100000 at USTCC.ust.edu.ph...
>
> Hi,
> Thanks for all those who replied.  I have a follow-up question. We are
> optimizing the use of 16SrDNA in our study. We are a little new to this
> endeavor, howver, when I run PCR on ATCC E.coli, the gel showed 2 bands:
> 1st band between 1330-1584 and the second band near 983. The Dna isolation
> i used is the genereleaser protocol.  Since I used the universal primers
> on this, I expect only the1st band.  Could the second band be a
> contaminant?
>
> Again, any advice is highly appreciated.
>
> Thanks.
>
> Sheila
>  *********************
>
> On Wed, 23 Feb 2000, Dr. Duncan Clark wrote:
>
> > In article <Pine.OSF.3.96.1000223143548.2605A-100000 at USTCC.ust.edu.ph>,
> > Ma Sheila M. De Jesus <mshmdeje at USTCC.ust.edu.ph> writes
> > >Does anyone know the meaning of the different bands in lambda DNA EcoRI
+ Hind
> > >III Marker from Promega
> >
> > Fragment sizes: 21226, 5148, 4973, 4277, 3530, 2027, 1904, 1584, 1330,
> > 983, 831, 564 and 125bp.
> >
> > Duncan
> >
> > --
> > The problem with being on the cutting edge is that you occasionally get
> > sliced from time to time....
> >
> > Duncan Clark
> > DNAmp Ltd.
> > Tel: +44(0)1252376288
> > FAX: +44(0)8701640382
> > http://www.dnamp.com
> > http://www.genesys.demon.co.uk
> >
>
> ---






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