Reverse Transcription - Subset of Jake’s O.lurida DNased RNA

Currently don’t have sufficient reagents to perform reverse transcription on the entire set of DNased RNA (control O.lurida ctenidia samples). To enable Jake to start testing out some of his primers while we wait for reagents to come in, Steven suggested I generate some cDNA for him to use.

Used the following DNased RNA:

• HC1

• NC1

• SC1

• HT1 1

• NT1 1

• ST1 1

Reverse Transcription Calcs: 20150522_Jake_Oly_cDNA_Calcs

Briefly:

• Reactions run in 0.5mL snap cap tubes

• 250ng of DNased RNA used in each reaction

• Combined DNased RNA with oligo dT primers and water; incubated 70C 5mins; immediately placed on ice

• Added 6.75μL of buffer/dNTP/enzyme master mix to each sample; incubated 42C for 1hr; 95C for 3mins

Samples will be given to Jake and stored @ -20C.