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Using mRNA in cloning - (Nov/09/2007 )

Hi All Scinetists Out There!

Having spent my possibilities using total RNA, I've now started to attempt cloning using mRNA extracted from gut tissue. However, having never used mRNA before, I'm a bit stuck on as to what concentration best used when running a PCR using cDNA made from mRNA.

Has anybody out there used mRNA succ essfully for cloning?


-Reasearch assistant-

My reverse transcription kit (Transcriptor, Roche) says use initialy 1ug of total RNA or 10 ng mRNA. (with limits 10 ng - 5 ug total RNA and 1 - 100 ng mRNA).