How degraded can an RNA sample be before amplification with RampUP?
If feasible, check the integrity of each RNA sample prior to amplification. If the RNA is intact, Random Primer (Vial 2) may be omitted from the first reverse transcription step (see protocol). If the RNA is partially degraded, use both random and dT primers in the first reverse transcription step (see protocol). The RampUP kits will amplify partially degraded RNA samples, but cannot amplify completely degraded RNA samples. One of the following methods should be used to determine the integrity of the sample: • Arcturus Paradise™ QC Process (Beta-Actin 3’/5’ value). Intact RNA samples have a QC Metric value between 1 and 10. Partially degraded RNA samples have a QC Metric value between 11 and 164. • Quantitation of ribosomal subunits. Intact RNA samples have a subunit ratio between 1.5 and 2.4. Partially degraded RNA samples have a subunit ratio less than 1.5. • Gel electrophoresis. Intact RNA samples have two sharp, bright bands of rRNA. Partially degraded RNA samples have some visible
