DNase Treatment - O.lurida Ctenidia 1hr Post-Mechanical Stress RNA

Quantified the RNA I isolated from Jake’s samples on 20150715 using the Roberts Lab NanoDrop1000 (ThermoFisher).

(http://eagle.fish.washington.edu/Arabidopsis/20150727_Oly_ctenidia_RNA_mech_stress_ODs.JPG)

(http://eagle.fish.washington.edu/Arabidopsis/20150727_Oly_ctenidia_RNA_mech_stress_plots_01.JPG)

(http://eagle.fish.washington.edu/Arabidopsis/20150727_Oly_ctenidia_RNA_mech_stress_plots_02.JPG)

Overall, the yields are good. The 260/280 ratios are mediocre. Will proceed with DNase treatment.

DNased 1.5ug of RNA from each sample using the Turbo DNA-free Kit (Ambion/Life Technologies), following the “rigorous” protocol.

Briefly:

• 50μL reactions were carried out in 0.5mL tubes

• added 1μL of DNase to each tube

• incubated 30mins @ 37C

• added additional 1μL of DNased

• incubated 30mins @ 37C

• added 0.2 vols (10.2μL) of DNase Inactivation Reagent

• incubated and mixed for 2mins @ RT

• spun 1.5mins, 10,000g @ RT

• transferred 50μL of supe to sterile 1.5mL snap cap tubes

• spec’d on Roberts Lab NanoDrop1000

Samples were stored @ -80C in Shellfish RNA Box #6. Will quantify at a later date.

DNase reaction calcs: 20150727_Jake_Oly_mech_stress_DNase_calcs