Setting up quantitative real-time PCR qRT-PCR

cDNA synthesis from RNA

✓ Reaction mix per sample

☐	5X RT buffer	4 µL
☐	dNTP-Mix	2 µL
☐	reverse transcriptase	1 µL
☐	random hexamer primer (160 µM), primer #312	1.25 µL
☐	oligo dT18 (100 µM); primer #945	0.1 µL
☐	RNA	1-2 µg
☐	RNA Inhibitor	0.5 µL
☐	–	–
☐	RNAse-free dd H2O	up to 20 µL

• Incubation of sample for 1 h at 37°C
• Heat-inactivation of enzyme at 70°C for 15 min

qRT-PCR

✓ Reaction mix per sample

☐	5X RT buffer	4 µL
☐	dNTP-Mix	2 µL
☐	reverse transcriptase	1 µL
☐	random hexamer primer (160 µM), primer #312	1.25 µL
☐	oligo dT18 (100 µM); primer #945	0.1 µL
☐	RNA	1-2 µg
☐	RNA Inhibitor	0.5 µL
☐	–	–
☐	RNAse-free dd H2O	up to 20 µL