Kaitlyn’s notebook: Geoduck hemolymph Reverse transcription (cDNA)

I made cDNA from samples I had successfully extracted RNA from (1, 2, 19, 27, 59, 62, 66, 11/15 Chewy, 11/15 Star, 11/21 Chewy, 11/21 Star).

Reverse transcription was performed using 50ng of each sample with M-MLV Reverse Transcriptase from Promega accroding to the Roberts Lab SOP. Calculations can be found here. A 1:1000 dilution of primer was made to increase pipetting volume.

Samples were initially heat to 45C for 5 min for denaturation instead of 70C, however the incubation was redone immediately after at 70C. This would not impact the cDNA because the step only serves to denature the RNA to maximize primer binding.

Samples are stored in the -20C fridge in 209 in 20190107 Kaitlyn’s Box. Additionally, the hemolymph cDNA created by Sam is also located in the same box.