CDNA synthesis

Use Biorad iScript cDNA Synthesis kit

Dilute RNA to 200ng/µl

Use 1µg of RNA for cDNA synthesis reaction

Prepare master mix (20µl reaction):
 * 5X Buffer 4µl
 * iScript Reverse Transcriptase 1µl
 * RNA (up to 1µg) 5µl (adjust accordingly)
 * H2O 10µl

Incubate at room temp for 5 min

Incubate at 42ºC for 1 hr

Stop the reaction by adding 100µl of H2O

Use 2µl of the diluted cDNA for each 20µl qPCR reaction