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?
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).