[Automated-sequencing] Re: Automated Sequencing of mRNA/cDNA

Phillip SanMiguel 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
> Sincerelly
>  
> Alex G.
> Gema Biotech sa
Hi Alex,
	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?
Phillip



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