Reverse Transcription - C.gigas RNA from Noah’s Heat-Mechanical Stress Project

reverse transcription
Crassostrea gigas
RNA
Pacific oyster
2023
Author

Sam White

Published

July 13, 2023

After isolating ctenidia RNA on 20230712 and then quantifying the RNA earlier today, I needed to perform reverse transcription to have cDNA for subsequent qPCRs. Reverse transcription was performed using oligo dT primers using M-MLV RT (Promega), per the manufacturer’s recommendations. Used 100ng of RNA in each reaction. All reactions were done on ice in 0.5uL PCR tubes.

In preparation for qPCR and primer testing, 2uL from each sample was pooled together in a separate 0.5uL tube. All cDNA was stored in Noah’s -80oC project box.