[Automated-sequencing] Re: Automated Sequencing of mRNA/cDNA
pmiguel at purdue.edu
Wed Aug 17 07:30:50 EST 2005
Alejandro Gonzalez wrote:
> Hi everybody.
> I'm working in automating sequencing of the plasmid and PCR templates,
> using MegaBACE 1000 DNA System Sequencer and the DyEnamic TM ET dye
> terminator Kit (GE-ex Amersham B.). I'd like to sequence by Sanger
> method a mRNA template or its cDNA, if anybody know how to do? or any
> tips to sequence this kind of template?, if anybody could send me a
> protocol to prepare this sequencing reaction and if I can use same kit
> (of GE-Amersham) I have been using.
> Thanks a lot
> Alex G.
> Gema Biotech sa
Your sequencing kit will not work for RNA because the kit uses a
DNA-templated DNA polymerase. However it should work fine for a cDNA.
Note that there must be only be the cDNA from one gene in your
sequencing reaction--if it is a mixture, sequencing the cDNA mixture
will not work.
What is the source of your mRNA?
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