4 replies to this topic

[oalkan]

Hi there,
does anyone has a real-time PCR protocol to quantify shRNA expression level driven from a U6 promoter? or can recommend a commercial kit?

[Functional Screens]

Just wondering, why do you want to do that?
Also, do you mean measuring the "mature" single strand siRNA?

[oalkan]

oalkan, on Mar 31 2010, 10:50 AM, said:

Hi there,
does anyone has a real-time PCR protocol to quantify shRNA expression level driven from a U6 promoter? or can recommend a commercial kit?

yes, I like to measure the the mature siRNA (processed shRNA). i wanted to compare the expression level under different promoter version. In additon, we made an inducible U6 promoter an dwe would like to measure th eiducer dependent expression of shRNAs

[oalkan]

Functional Screens, on Mar 31 2010, 11:05 PM, said:

Just wondering, why do you want to do that?
Also, do you mean measuring the "mature" single strand siRNA?

yes, I like to measure the the mature siRNA (processed shRNA). i wanted to compare the expression level under different promoter version. In additon, we made an inducible U6 promoter an dwe would like to measure th eiducer dependent expression of shRNAs

[Functional Screens]

To make it easy, why don't you express a short irrelevant sequence instead of shRNA sequence since your purpose is to measure the expression level of transcript under induction (or different promoters).