Re: Autoseq Digest, Vol 38, Issue 2
David Reynolds
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 (dna...@flinders.edu.au)
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>----------------------------------------------------------------------
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>Message: 1
>Date: Tue, 19 May 2009 23:44:33 +0930
>From: dna...@flinders.edu.au
>Subject: [Automated-sequencing] injection solution which reduces rfu
>To: aut...@magpie.bio.indiana.edu
>Message-ID: <1242742473.4...@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: drey...@aecom.yu.edu
http://www.aecom.yu.edu/dna/
________________________________________________________________