Table 1.

Reverse transcription reaction mix

Component	Stock concentration	Final concentration	Amount per sample
RNase-free water	n/a	n/a	To 20 μL
Total RNA	n/a	n/a	1 μga (Volume may vary among samples)
cDNA Primer Mix	20 μM	20 pmol	1 μL
SCRIPTUM FIRST RT buffer	5×	1×	4 μL
dNTP Mix	10 mM per dNTP	500 nM per dNTP	1 μL
DTT	100 mM	5 mM	1 μL
RNase inhibitor	40 units/μL	40 units	1 μL
SCRIPTUM FIRST Reverse Transcriptase	200 units/μL	100 units	0.5 μL
Total	n/a	n/a	20 μL

^aAccording to the manufacturer’s protocol, the amount of starting material can vary from 1 pg - 5 μg of total RNA. Therefore, also lower RNA amounts are sufficient. For example, we have successfully used about 0.1–0.3 μg of RNA.