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PCR product as template for in vitro transcription

In vitro transcription PCR RNA Probe In situ hybridisation

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#1 Sugan

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Posted 30 June 2013 - 09:14 PM

I am searching for options of templates for PCR reaction, to use the resultant PCR product as a template for in vitro transcription. I came through options such as plasmid containing gene of interest (GOI) and genomic DNA. Can I use first strand cDNA as a template for PCR amplification with primers (containing the T7 promoter sequence) specific to GOI ? Will this give a product incorporated with T7 promoter sequence which I can use for in vitro transcription?

#2 bob1

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Posted 01 July 2013 - 12:58 PM

Potentially yes, but it would be easier and quicker in the long run to clone it into a vector with a T7 promoter built in. That way you wouldn't run into the problem of having errors in the sequence from the PCR and would be able to make any amount of the plasmid you wished with no difficulties.





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