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ARS Home » Midwest Area » Madison, Wisconsin » Vegetable Crops Research » Research » Publications at this Location » Publication #242452
Title: Effect of RNA Integrity Determined With the Agilent 2100 Bioanalyzer on Bacterial RNA Quantification with RT-PCR

Author
item JAHN, COURTNEY - University Of Wisconsin
item CHARKOWSKI, AMY - University Of Wisconsin
item Willis, David

Submitted to: Annual Quantitative Polymerase Chain Reaction (PCR) Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 9/19/2008
Publication Date: 3/16/2009
Citation: Jahn, C.E., Charkowski, A.O., Willis, D.K. 2009. Effect of RNA Integrity Determined With the Agilent 2100 Bioanalyzer on Bacterial RNA Quantification with RT-PCR [abstract]. Annual Quantitative Polymerase Chain Reaction (PCR) Meeting.

Interpretive Summary:

Technical Abstract: RNA integrity is critical for successful RNA quantification. The level of integrity required differs among sources and extraction procedures and has not been determined for bacterial RNA. Three RNA isolation methods were evaluated for their ability to produce high quality RNA from D. dadantii. The isolation methods differed significantly in yield, RNA quality, as determined by the RNA Integrity Number (RIN) with the Agilent 2100 bioanalyzer, and amount of contaminating DNA. Furthermore the influence of RNA quality on the outcome of real-time qRT-PCR was studied. The assessment of RNA integrity was critical for obtaining meaningful gene expression data. RIN values below 7 resulted in high variation and loss of statistical significance.