[Automated-sequencing] Re: Autoseq Digest, Vol 38, Issue 2

David Reynolds via autoseq%40net.bio.net (by dreynold from aecom.yu.edu)
Tue May 19 18:11:09 EST 2009


Hi Oliver,
Instead of, or in conjuction with an additive you can dilute the 
fragments to get reasonable RFUs.  On our 3730 the PCR reactions were 
typically diluted 40 to 120 fold.  We would titrate a few samples per 
marker at 40, 80 and 120 fold, then pick the best dilution for the 
rest of the study.
Regards,
Dave

At 01:03 PM 5/19/2009, you wrote:
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>    1. injection solution which reduces rfu (dna0001 from flinders.edu.au)
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>----------------------------------------------------------------------
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>Message: 1
>Date: Tue, 19 May 2009 23:44:33 +0930
>From: dna0001 from flinders.edu.au
>Subject: [Automated-sequencing] injection solution which reduces rfu
>To: autoseq from magpie.bio.indiana.edu
>Message-ID: <1242742473.4a12bec9391eb from imp.flinders.edu.au>
>Content-Type: text/plain; charset=ISO-8859-1
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>
>
>Hi all
>     I work in a core lab setting, and have recently upgraded my Applied
>Biosystems 3100 Genetic Analyser to a 3130xl running POP-7. A client wants to
>perform fragment analysis with multiplexed PCR products, but often 
>the peaks of
>interest are off-scale (i.e. > 7000 rfu). To the best of my knowledge, Gene
>Mapper v4.0 and Peak Scanner v1.0 will not analyse these samples due to the
>off-scale peaks. I've tried adding 0.1mM EDTA to the samples (we generally use
>HiDi formamide only for injection), but have found that this can either
>increase or decrease signal strength. Can anyone suggest a solution which will
>decrease signal strength? Will low melting point agarose achieve this? Any
>suggestions would be appreciated.
>
>Many thanks
>
>Oliver
>
>
>
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>End of Autoseq Digest, Vol 38, Issue 2
>**************************************

________________________________________________________________

  David Reynolds
  Director
  Genomics & Genetics Core
  Albert Einstein College of Medicine
  Price Center for Genetic and Translational Medicine
  Room 419
  1301 Morris Park Avenue
  Bronx, New York 10461-1602

  Tel: (718) 678-1157
  Lab: (718) 678-1160
  Fax: (718) 678-1016
  E-mail: dreynold from aecom.yu.edu
http://www.aecom.yu.edu/dna/

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