My problem is that I obtained 18s hostgene expression appears on very late cycles. cDNA quality may be low. OD260/280:1.68 and 640ng/ul
500ng RNA+ NF water to final volum 10ul is warmed 70 C for 5 min,
Then added 10ul Mastermix to RNA (total volum 20ul) and incubate it 42 C for 1 hour and 94 C 2 minute andd 4 C forewer.
Mastermix:
Finally 40 ul water is added to tubes. cDNA ready.