Currently don’t have sufficient reagents to perform reverse transcription on the entire set of DNased RNA (control and 1hr.heat-shocked 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.