All of the RNA (50uL) was DNase treated with Ambion’s Turbo DNA-free kit according to protocol. Samples were spec’d.
Results:
Tag Archives: DNase
gDNA Removal – Abalone RNA from 20090402 and 20090331
Transferred 50uL of RNA to fresh tubes and processed them using the Ambion Turbo DNA-free kit according to the manufacturer’s protocol.
RNA Precipitation – Hard clam gill #1 RNA from 20080819
DNAsed RNA using Ambion Turbo DNA-free kit, following the rigorous procedure. Diluted total RNA to 0.2ug/uL (Vf = 720uL). Added 1uL DNase and incubated the tube @ 37C for 30mins. Added an additional 1uL DNase and continued incubated for 30mins. Added 0.2 volumes of DNase Inactivation Reagent (158.4uL) and incubated at RT for 10mins with periodic mixing. Pelleted inactivation reagent according to protocol and transferred supe (DNA-free RNA) to clean tube.
Results: RNA looks really nice. Have a large quantity of RNA (700uL x 0.2275ug/uL = 159.25ug). Will split into four equal parts and isolate mRNA from 3 of the 4. Those three will be:
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Ambion kit x 1
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Ambion kit x 2
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Promega PolyA Tract kit.
RNA Isolation – V.tubiashii samples from autoclaved gigas exposure (from 20081218)
RNA was isolated from the Control and V.tub+gigas samples from the 0, 1, & 24hr time points using 1mL TriReagent. No visible pellets. Used 20uL of 0.1%DEPC-H2O to resuspend RNA. Incubated @ 55C, 5mins. Spec’d.
Results: RNA looks OK, but not great. For the “V.tub + gigas t=1″ sample, the third spec reading is correct. The first two had the air bubble error.
DNAse Treatment – V.tubiashii total RNA (see above)
1ug of RNA in a volume of 12uL was DNAsed using the Ambion DNA-free Kit according to their protocol. RNA was transferred to a fresh tube and stored @ -80C in Sam’s RNA Box #1.