qPCR – Withering Syndrome RNA Check

Ran withering syndrome qPCR on the following RNA samples provided by Lisa to determine if it’s necessary to DNase treat them prior to performing reverse transcription:

I believe these were extracted from water filters collected from basement abalone tank water. The OD260/280 ratios are pretty bad, particularly for RNA, but will proceed anyway.

Master mix calcs are here: 20150318 – qPCR WSN RNA Test

Since I’m anticipating using 1000ng of RNA in the reverse transcription reactions (25μL reaction volume = 40ng/μL), I only used 1μL of RNA (~100ng), and positive control, template in each reaction to semi-accurately represent the quantity of RNA that would be loaded in a qPCR reaction with 2μL of cDNA.

All samples were run in duplicate. Positive control was the 3e6 p18RK7 standard curve sample from 20120731.

Results:

qPCR Report (PDF): Sam_2015-03-18 10-11-35_CC009827.pdf
qPCR Data File (CFX96): Sam_2015-03-18 10-11-35_CC009827.pcrd

All samples are positive for withering syndrome. Will proceed with DNase treatment.

2 thoughts on “qPCR – Withering Syndrome RNA Check

  1. Pingback: qPCR – Withering Syndrome cDNA Tests | Sam's Notebook

  2. Pingback: DNase – Withering Syndrome RNA | Sam's Notebook

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