URGENT! RAPD vs. AFLP among vampire bats

Scott Handley ssh4 at cdc.gov
Tue Aug 24 12:26:02 EST 1999


Good luck with the AFLP kit.  PE's kit comes with its share of problmes.  I would
suggest designing your own primers and adaptors, and then just use your own
in-house reagents.  There are many many papers out there to use as references for
this.  Plus I would look into IRS-PCR (infrequent-restriction-site PCR).  This is a
novel technique for doing AFLP type experiments.  I have been doing AFLP, RAPD's
and IRS-PCR for about a year now and the most consistent method is IRS-PCR.
However, this is on bacteria ... not vampire bats.  But DNA is DNA.


Anne Gillen wrote:

> Hello,
>
> I have had trouble repeating RAPD markers.  I would not rely on them for
> fingerprinting.  I have tried to repeat markers developed in other labs
> using the same cultivar and I have not been able to repeat them.  DNA
> quality, thermal cycle parameters and brand of thermalcycler all affect
> RAPDs. However, I have found that some bands are very repeatable within my
> lab.
>
> I have had trouble getting my AFLP digest-ligations to always work
> so am switching to the kit in hopes of more consistent results. I cannot
> say from experience, but it seems that AFLPs are much better than RAPDS
> for repeatability.
>
> *******************************************************************************
>     Anne M. Gillen                           E-mail: amgillen at ucdavis.edu
>     Pomology Department                      Phone:  (530) 752-1462
>     University of California at Davis
> *******************************************************************************




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