M13 primers and mtDNA sequencing

Krzysztof Lubieniecki klub at agrpc209.ar.lublin.pl
Tue Mar 21 15:40:49 EST 2000


Dear Sequencers,

I have just started with sequencing D-loop region of mtDNA. I will be
using Big Dye Terminator cycle sequencing kit. I am expecting a
product of about 900 bp. I was planning to order  regular primers but
I have heard that conditions for the sequencing reaction given in the
kit where optimized for the primers tailed with -21 M13 forward
primer and therefore the profile given in the kit may not work for
regular primers (untailed)and it may be necessary to optimize the
sequencing reaction which is supposedly difficult to do.
Has anyone heard about it or has any comments about it?

         Krzysztof


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Krzysztof Lubieniecki 
Department of Immunogenetics
Institute of Genetics and Animal Breeding
Polish Academy of Sciences, Jastrzebiec
05-551 Mrokow
Fax: +48-22-7561699
Phone: +48-22-7561711
email: klub at agrpc209.ar.lublin.pl
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