What testing should be performed to assess the quality of an RNA sample?
FAQ ID -2660

All RNA samples should be assessed spectrophotometrically (diluted in 10mM Tris, pH 8.0), and electrophoretically, and should meet the following specifications:

  • Total RNA concentration by A260 should be greater than 40 µg/ml
  • A260: A280 ratio should be 1.8 to 2.0
  • A260: A230 ratio should be greater than 1.7
  • Analysis of ~100ng of total RNA on an Agilent Bioanalyzer using an RNA 6000 Nano LabChip, or analysis of 1.5 μg of total RNA on a denaturing 2.0% agarose gel containing ethidium bromide (0.5 μg/ml) should contain sharp 28S and 18S rRNA bands, with no smearing at their low molecular weight edge. The 28S:18S band intensity ratio should be ~2:1. When utilizing the RNA 6000 Nano LabChip for RNA analysis, the RNA should have a RIN (RNA integrity) score of 7.0 or higher.

In addition to the above quality control tests, the RT2 RNA QC PCR Array for human (PAHS-999), mouse (PAMM-999), or rat (PARN-999) can be used. These arrays allow the rapid assessment of high and low housekeeping gene expression levels, reverse transcription and polymerase chain reaction efficiency, and genomic and general DNA contamination.