I extracted ~87 ug of total RNA from mushroom samples using TRIzol (Invitrogen) and I got ~1,2 ug of poly-A RNA using the Oligotex kit (QIAGEN) for mRNA purification. The protocol of direct cDNA sequencing kit (SQK-DCS109) recommends to start with 100 ng of poly-A RNA. I repeated this protocol twice and when I quantified the samples at the end of the second strand synthesis step using QUBIT (dsDNA high sensitivity kit), the amount is too low.

Should I start the cDNA synthesis using more than 100 ng of poly-A RNA?

Someone has already done it?