Dear All, 1 ug of permissible quality FFPE RNA is transcribed to cDNA through SSIV VILO RT kit.

I have Qubit v.4 device, which can measure cDNA to a good extent. The cDNA ranges from 25-100 ng / ul for these FFPE samples through Qubit. Now, is there any rule regarding cDNA input to create libraries for RNA SEQ to be performed through Ion Torrent platform?

For FFPE samples, we always use more template to create libraries (like in whole exome or whole genome sequencing), but for cDNA library I am not much sure that what should be the starting cDNA amount to get 100 pM range libraries. If I make libraries with starting material 50-60 ng of total cDNA, would it be suffice, if my samples are FFPEs in origin?

Also, if someone does not have Qubit to measure cDNA, so what strategy we should use for the cDNA starting quantity for RNA SEQ library preparations after reverse transcribing the RNA?

Thanks....