Degradation of RNA can interfere with measurement of RNA expression either by making a sample too degraded to be analysable or by distorting the result which is obtained. RNA degradation is commonly assessed by electrophoretic analysis but this technique is qualitative rather than quantitative. Brisco and Morley recently described a simple PCR technique whereby several amplicons of a reference gene were quantified to obtain a quantitative measure of RNA degradation. The measured transcript number could then be corrected for degradation by using this measure of degradation together with the length of the test amplicon being studied. The technique is simple, gives a quantitative result and only requires 1-50 pg of RNA, depending on the extent of degradation.
Monoquant provides a kit suitable for quantification of degradation in samples of human RNA. The reference gene is glyceraldehyde phosphate dehydrogenase.
For further details please contact Professor Alec Morely at firstname.lastname@example.org.
Brisco, M.J and Morley, A.A. Quantification of RNA integrity and its use for measurement of transcript number. Nucleic Acids Research 2012; doi: 10.1093/nar/gks588