Cdna synthesis using non-random primers - Google …
Both the pd(N)6 (80 nmol) and pd(N)9 (50 nmol) random primers contain approximately 150 µg (5 OD) and are ideal for use in cDNA synthesis, RT-PCR and hybridization applications.
(12S or 16S) in cDNA amplified using random primers (N7) ..
We are running a real time PCR experiment to study and compare the expression of mRNA in different treatment groups. We have been running our reverse transcription using random hex on the recommendation of a neighboring lab. We then used that synthesized cDNA in our real time PCR runs. While comparison of total RNA is valuable for our study, we are more interested in mRNA expression at the moment. Should we be running our reverse transcription using oligo dt? What is confusing me is that in the literature we used as a guideline for our work, they claim to be studying mRNA expression, while running the reverse transcription using random hexs. Any input would be helpful, thanks for the help.
(I assume that the rRNA bands will be relatively less bright when using oligo-dT primers than when using the random primers, since the latter also creates cDNA from the rRNA.)
Random hexamers also generate shorter cDNAs, especially if their concentration is high, so the shape of the smears might be different.
Can I ask why you are so preoccupied with verifying the products of your RT?