View source for Setting up quantitative real-time PCR qRT-PCR

== cDNA synthesis from RNA ==
{{Pipetten-Checklist-8step
|schritt1=5X RT buffer|menge1=4 µL

|schritt3=reverse transcriptase|menge3=1 µL|menge2=2 µL|schritt2=dNTP-Mix|schritt4=random hexamer primer (160 µM), primer #312|menge4=1.25 µL|schritt5=oligo dT18 (100 µM); primer #945|menge5=0.1 µL|schritt6=RNA|menge6=1-2 µg|schritt7=RNA Inhibitor|menge7=0.5 µL|schritt8=–|menge8=–|zusatz1=RNAse-free dd H2O|titel=Reaction mix per sample|gesamt=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 ==
{{Pipetten-Checklist-8step
|schritt1=5X RT buffer|menge1=4 µL

|schritt3=reverse transcriptase|menge3=1 µL|menge2=2 µL|schritt2=dNTP-Mix|schritt4=random hexamer primer (160 µM), primer #312|menge4=1.25 µL|schritt5=oligo dT18 (100 µM); primer #945|menge5=0.1 µL|schritt6=RNA|menge6=1-2 µg|schritt7=RNA Inhibitor|menge7=0.5 µL|schritt8=–|menge8=–|zusatz1=RNAse-free dd H2O|titel=Reaction mix per sample|gesamt=up to 20 µL}}