RNAse protection assay - efficient for a high number of probes ?

Gary Kucera kucera~gt at glaxowellcome.com
Wed Oct 29 08:34:23 EST 1997


jzuber at hotmail.com wrote:
> 
> We are working with subtractive cDNA-Libraries. To check the
> differentiality of our subtracted clones we used to use Northern
> blotting. As we get to know now this is a quite time consuming procedure,
> because the number of potentially differential and interesting clones is
> exploding day by day.
> 
> RPA is supposed to be at least as sensitive as Northern blotting. Do you
> know any protocol/kit/procedure to use RPA for a high number of probes
> parallely (we want to check all in all 300 clones per library) and what is
> your experience about it ?
> 
> Thanks&Bye,
> Hannes
> 
> -------------------==== Posted via Deja News ====-----------------------
>       http://www.dejanews.com/     Search, Read, Post to Usenet



With that many samples, and considering you want to check numerous
probes, you may want to consider Quantitative RT-PCR.  In response to
your original question, I have used Ambion's RPA kit with very little
success and instead use a "homemade" protocol with excellent results. 
If you are interested I can send you the protocol.  But consider the
Quant. RT-PCR option.



More information about the Methods mailing list