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From what Ive seen of taqman. (presuming the same number of cycles between your gel and PCR) it could be that the 24mer is a bit too long for use as a probe. Ive been told that 20mer probes are bordering on too long. Its sadly one of the problems with Taqman.
SYBR green in my experience is only a little less accurate than Taqman. And if you can show specificity by PCR gel (and maybe northern blot if you wanted to be anal about it), its generally well accepted.
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In Topic: One siRNA sequence, many cell lines
14 June 2012 - 06:03 AM
jmafs, on 13 June 2012 - 09:41 AM, said:
Hi all,
I'm new to process of siRNA transfection and I was wondering: Will one siRNA sequence (previously validated in the lab) be good enough to transfect multiple other cell lines from the same organism? I understand that the actual process of transfection will be different for each cell line, but I am curious as to whether I need to also worry about the sequence itself.
Thanks!
I'm new to process of siRNA transfection and I was wondering: Will one siRNA sequence (previously validated in the lab) be good enough to transfect multiple other cell lines from the same organism? I understand that the actual process of transfection will be different for each cell line, but I am curious as to whether I need to also worry about the sequence itself.
Thanks!
Sequence shouldnt be a problem for transient or stable transfections tbh, you may need to change the vector but apart from that the sequence should not change.
Depending on the gene its knockdown could kill the cell,though tbh, that is a rare occurance and is usually only if its a cell signalling protien key to anti-apoptotic signalling
