Withering syndrome RNA, extracted from water filters, was confirmed via qPCR to contain detectable quantities of withering syndrome DNA. Before proceeding to make cDNA, residual gDNA carryover from the RNA extraction needs to be removed.
Performed DNase treatment with the Turbo DNA-free Kit (Ambion/Life Technologies), following the rigorous. Used 10μL of each of the following template RNA:
- Day 0-1
- Day 3-1
- Day 7-1
- Day 11-1
Reactions were performed in a volume of 50μL. Used 1μL of DNase for the first 30mins @ 37C and then added an additional 1μL of DNase for the final 30mins @ 37C. Samples were inactivated according to the manufacturer’s protocol and spec’d on the Roberts Lab NanoDrop1000.
Yields are consistent. The OD260/280 values are still poor (didn’t expect them to change, though). Will qPCR to verify removal of gDNA.