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shRNA expression levels


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4 replies to this topic

#1 oalkan

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Posted 31 March 2010 - 06:50 AM

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?

#2 Functional Screens

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Posted 31 March 2010 - 07:05 PM

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

#3 oalkan

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Posted 01 April 2010 - 10:41 AM

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

#4 oalkan

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Posted 01 April 2010 - 10:42 AM

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

#5 Functional Screens

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Posted 01 April 2010 - 04:06 PM

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).




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