internal control in semi quantitative rt pcr - question (Jun/10/2005 )
I want to evaluate the expression of low copy number transcript by semi quantitative RT-PCR approach . I have an internal control . can I do the pcr reactions of this 2 genes in 2 seperate tubes ( to prevent the interference of one gene amplifiication on the other ) or they "must" coamplified in multiplex pcr manner to get a semi quantitative results ?
Either way is OK and acceptable for publication.
Both one tube and seperate tube is ok.
But you have to make sure both of your primer sets have the similar PCR efficiency before you can make comparison.
You primer efficiency can be done by software prediction.