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