control for cDNA synthesis - (Apr/21/2008 )

Hi,

What do you guys use as control for cDNA synthesis?
(a) water, reagents & enzymes that synthesize cDNA (minus RNA)
( RNA without RT
or any others?

Thanks in advance:)

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I just use -RT as a control (a -RT reaction for each sample): RNA and all other reagents except RT enzyme -- using water as replacement for enzyme.

Then in PCR, I use primers for a housekeeping gene as a positive control for the +RT's, which also shows (by absence of a band, hopefully ) no contamination in -RT. Also, always run a NT (no template) control PCR reaction.

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also a -RT control. another possibility you have is to design your primers in regions that contain introns. that way even if you have DNA contamination you'll have a different size in bands, the cDNA will have a smaller size than the band correspoding to the contamination.
cheers

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