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[Zbrafish] WMISH - RNA probe synthesis - doubt

Nagganatha Suganthan R via zbrafish%40net.bio.net (by btsugan from gmail.com)
Sun Jul 7 09:52:54 EST 2013


Dear members

I have a question regarding the primer design for amplifying probe
templates containing an RNA polymerase promoter to produce RNA probe.

While designing the primers attached with T7 promoter sequence, just adding
the promoter sequence at 5' extremity of the primer is enough or a short
sequence of GAAAT should also be added before the promoter sequence? Is it
either 5'-TAATACGACTCACTATAGGGnnnnnnn-3' or
5'-gaatTAATACGACTCACTATAGGGnnnnnnn-3'? The template to be used for the PCR
is cDNA.

Eagerly waiting for a reply

Thanks and Regards

Suganthan
PhD Scholar
Ehime University
Japan
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