Assessing integrity of insect RNA

Authors: Fabrick, Jeffrey; Hull, Joe

Submitted to: USDA-ARS Research Notes
Publication Type: Research Notes
Publication Acceptance Date: 3/8/2017
Publication Date: 4/12/2017
Citation: Fabrick, J.A., Hull, J.J. 2017. Assessing integrity of insect RNA. USDA-ARS Research Notes. 5991-7903EN: 1-12.

Interpretive Summary: Obtaining high-quality genetic material (DNA or RNA) from biological samples is necessary for downstream molecular applications, including amplification and sequencing protocols. Hence, having appropriate tools to evaluate the quality and integrity of such material is essential. Currently, capillary electrophoresis instruments are commonly used to assess the quality of RNA/DNA from most biological sources. While the instrumentation is often optimized for genetic material from well-characterized species, these model species are frequently limited to vertebrates. Many non-model organisms, however, have unique and highly complex RNA profiles that obstruct the interpretation of RNA quality. Here, ARS scientists at Maricopa, AZ show that considerable differences in insect RNA profiles can impede quality assessments and that degraded RNA encumbers the outcome of downstream molecular applications. The results reveal that although insect RNA profiles differ; nonetheless, commercially-available capillary electrophoresis instrumentation can be used to assess the quality of insect RNA profiles.

Technical Abstract: Assessing total RNA integrity is important for the success of downstream RNA applications. The 2100 Bioanalyzer system with the RNA Integrity Number (RIN) provides a quantitative measure of RNA degradation. Although RINs may not be ascertained for RNA from all organisms, namely those with unusual or complex RNA profiles, this application note demonstrates the applicability of total RNA quality assessment of different insect species with the 2100 Bioanalyzer system and its relevance to downstream quantitative RT-PCR experiments.