Measuring cDNA concentration - (May/29/2008 )
Hi-
Is it necessary to measure the concentration of cDNA for a gene expression qPCR experiment, if RNA concentration is measured before reverse transcription? In all of the published work I've referenced, only RNA concentration was measured (before reverse transcription), but I've never seen cDNA concentration being measured. However, I can see the merit in measuring cDNA concentration before qPCR, which is why I'm wondering if it is necessary. Thanks.
-sgt_pepper06-
QUOTE (sgt_pepper06 @ May 29 2008, 10:54 AM)
Hi-
Is it necessary to measure the concentration of cDNA for a gene expression qPCR experiment, if RNA concentration is measured before reverse transcription? In all of the published work I've referenced, only RNA concentration was measured (before reverse transcription), but I've never seen cDNA concentration being measured. However, I can see the merit in measuring cDNA concentration before qPCR, which is why I'm wondering if it is necessary. Thanks.
Is it necessary to measure the concentration of cDNA for a gene expression qPCR experiment, if RNA concentration is measured before reverse transcription? In all of the published work I've referenced, only RNA concentration was measured (before reverse transcription), but I've never seen cDNA concentration being measured. However, I can see the merit in measuring cDNA concentration before qPCR, which is why I'm wondering if it is necessary. Thanks.
It is not necessary.
-cellcounter-
QUOTE (cellcounter @ May 29 2008, 02:28 PM)
It is not necessary.
Yes, no need to measure.
I thing one important reason is that isolate cDNA along is too much of work.
Just make sure those samples you will run together do RT together, so that the variation of RT efficient among samples will be as small as possible.
-wuxx0153-