Quantified the RNA I isolated from Jake’s samples on 20150715 and 20150710 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:
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50μL reactions were carried out in 0.5mL tubes
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added 1μL of DNase to each tube
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incubated 30mins @ 37C
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added additional 1μL of DNased
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incubated 30mins @ 37C
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added 0.2 vols (10.2μL) of DNase Inactivation Reagent
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incubated and mixed for 2mins @ RT
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spun 1.5mins, 10,000g @ RT
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transferred 50μL of supe to sterile 1.5mL snap cap tubes
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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