ABI sequencing artifact

dina at libpc1.icos.com dina at libpc1.icos.com
Fri Sep 16 18:43:03 EST 1994


In article <quinnt-1409941902420001 at microb3.biostat.washington.edu>, 
<quinnt at u.washington.edu> writes:
> Path: 
news.halcyon.com!nwnexus!scipio.cyberstore.ca!nntp.cs.ubc.ca!news.bc.net!news.m
ic.ucla.edu!library.ucla.edu!csulb.edu!nic-nac.CSU.net!usc!howland.reston.ans.n
et!vixen.cso.uiuc.edu!news.uoregon.edu!netnews.nwnet.net!news.u.washington.edu!
microb3.biostat.washington.edu!user
> From: quinnt at u.washington.edu (Matthew Stoecker)
> Newsgroups: bionet.molbio.methds-reagnts
> Subject: ABI sequencing artifact
> Date: Wed, 14 Sep 1994 19:02:42 -0800
> Organization: University of Washington
> Lines: 10
> Message-ID: <quinnt-1409941902420001 at microb3.biostat.washington.edu>
> NNTP-Posting-Host: microb3.biostat.washington.edu
> 
> I have been using the ABI DyeDeoxy kit with Taq for sequencing and have
> encountered a strange artifact.  On one particular template, I get
> beautiful read for about 200 bp, then spotty read for about 10 bp, then
> absolutely nothing for the rest of the run.  This has happened twice, and
> the break always occurs in the same spot.  I was wondering if anyone had
> any similar occurences or any ideas about what's going on.  Thanks a
> lot!!!
> 
> Matthew Stoecker
> U. of Washington

The only times I have seen this happen is:
1) If there is too much DNA in the sequencing reaction
2) If you are sequencing a PCR product

I usually find that it is an overdose of DNA in the reaction mix and all the 
dyes are taken up in the first few rounds of the cycle sequencing PCR.



More information about the Methods mailing list