The Clontech RNA/cDNA Quality Assay Kit directly determines the quality of your human RNA and cDNA samples using reverse transcription (RT) and PCR. To ensure the representation of full-length transcripts for experiments with cDNA libraries, RACE-ready cDNAs, or cDNA microarray probes, use the Clontech RNA/cDNA Quality Assay to check the integrity of your RNA starting material before beginning your experiment. Because this assay uses RT-PCR, it provides a direct functional test of your sample for its ability to produce full-length cDNA for your application. You achieve quick results using standard lab equipment, and avoid inconvenient and toxic formaldehyde gels.
Analyze RNA and cDNA Integrity
While RNA can look intact on a formaldehyde gel or in a chip-based assay (28S:18S ratio >1), RNA can be degraded during cDNA synthesis. The result: first-strand cDNA containing a mix of truncated and full-length transcripts.
Using our RNA/cDNA Quality Assay ensures that you won’t waste time or get invalid results by using low-quality RNA in demanding applications. The assay uses two sets of primers in a multiplex RT-PCR to amplify 5'- and 3'-end fragments of a long mRNA (~3 kb) for a carefully selected housekeeping gene. Since in most cases RNA degradation starts in the 5'-end region of an RNA molecule, the ratio of the 3' to 5' amplified fragments provides a direct indication of RNA integrity.