advice needed for check trace mRNA

Martin Leach leach at mbcrr.harvard.edu
Wed Apr 20 10:16:30 EST 1994


In article <1994Apr18.151548.1 at leif>, ldeng at kean.ucs.mun.ca (GEORGE XIAOXI
OUYANG, ELECTRICAL ENGINEERING, PHONE 579-0970) wrote:

> Hi! Dear Netters! I want to identify very little trace of mRNA in the cells 
> by RT-PCR or Northern hybridization. I certainly want to show the RT-PCR 
> signal is not come from DNA. So anyone can tell me what is the best way to 
> extract RNA which is free of DNA?
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^

Virtually impossible!


 What is the best probe (for Lymphocytes) 
> for the very-low-expression mRNA in Northern hybridization? Thanks in 
> advance!  Langtuo Deng at: ldeng at kean.ucs.mun.ca

1. Either try RNAse protection - more sensitive than Northerns.

2. Or try RT-pcr with primers that span at least one intron. - so if DNA
contamination it either will be larger product or Un-PCRable if too big.

Martin Leach

-- 

.....          Martin Leach                Email:leach at mbcrr.harvard.edu 
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