Considerations For Extraction, Stabilization And Quantification By PCR For Sensitive Sample Such As RNA

This article talks about careful selection of an extraction system that helps stabilize and protect targets, especially RNA, or which allows for immediate analysis.

The accessibility of PCR and qPCR systems has increased to where they are almost considered a commodity product today. This is certainly the case for end-point PCR systems and some qPCR units also allowing researchers to carry out gene expression analysis more conveniently and quickly. This accessibility has been an innovative development in the community and will continue to move research forward for years to come.

There have been concerns with regards to reverse transcriptase (RT-PCR) and real-time RT-PCR analysis of sensitive sample types such as RNA due to the stability of the sample during process and storage. Just how much impact this has had upon the results may go unknown. Recent advances into techniques such as digital PCR and new advances in extraction stabilization systems have sought to address some of these concerns.

Extraction

In theory, the extraction process is a relatively trivial matter: disrupt the sample’s cellular integrity and collect the released nucleic acids. However, it is never that simple. Multiple factors must be addressed.

When performing any nucleic acid extraction (especially RNA), stability and integrity of that extract must be considered. Samples being extracted in remote field studies must also take into consideration logistical concerns such as power supply, temperature control, availability to equipment, and shipping if samples are not to be tested onsite immediately.