Quantitative RT-PCR (addendum)

brett brett at BORCIM.WUSTL.EDU
Tue Oct 14 09:38:41 EST 1997


>>Hello,
>>We would like quantitate the level of gene expression by means of RT-PCR
>>(Northern cannot be used since very limited amount of RNA is available).
>>Anybody who is experienced in doing quantitative RT-PCR please kindly
>>e-mail me the protocols. By the way, would RNase protection assay be a
>>good alternative?
>>Many thanks.
>>Alan Lee
>>e-mail: ywlee at hkucc.hku.hk
>
>RT-PCR would be good, although you'd be wise to optimize and work out your
>controls on in-vitro transcribed RNA. RPA is also good. I was able to
>quantitate
>a viral message down to 1 copy per cell with a really hot probe.  I can supply
>you with protocols if you wish.
>
just to clarify, that is 1 copy/cell from 106 cells (ie quantitative detection
limit ~106)! sorry.


Brett Lindenbach
                             
Washington University - St Louis                  
brett at borcim.wustl.edu                             




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