1 reply to this topic

[smjazayeri]

I am using GeneAmp kit in order to convert mRNA to cDNA but the kit says that after conversion mRNA to cDNA it is possible to amplify the cDNA with primers as gene specific or other primers. So as I have not got too much of the samples I want to amplify cDNA before doing RT-PCR. I would like to know that it is possible to amplify the cDNA with hexamer or oligo dT primers in order to have more and more copies of the genes or what?
Thanks
SMJ

[kutayozturk]

In order to amplify cDNAs and then use in qRT-PCR analysis, I think you can use SMART RACE. I don't know how GeneAmp kit works. However, even very low amount of cDNA is enough for qRT-PCR. I have obtained bands from cDNAs derived from 39ng mRNA (you may need some more for total RNA), even if I dilute the cDNA 50 times. In addition, qRT-PCR is much more sensitive than conventional PCR, so I don't think you really need to do amplification