Ensure High Quantity And Quality Of Isolated Bacterial Ribonucleic Acid (RNA)

By Jaydeep Chaudhuri and Nadia Kadi, Pall Corporation

Molecular exploration in bacteria has undergone a substantial change over the past decade. Emerging evidences indicate that the complex signal coordination around riboswitches, RNA thermometers and small bacterial RNAs is regulated by RNAs. Moreover, the growing interest in human host-pathogens interactions has fast-tracked the transition of metagenomic next generation sequencing (mNGS) from a research tool into a valuable clinical tool. In all these cases the journey begins with high quality RNA.

The quantity and quality of the input RNA determines the accuracy and reproducibility of an experiment. Depending on the demand for the quality of the input RNA, either a spectrophotometric, fluorometric, or capillary electrophoresis is performed to assess quality control on input nucleic acids. RNA is extremely susceptible to degradation because of the omnipresent RNase in the environment and precautions must be taken for successful extraction of high-quality RNA.

In this scientific brief we evaluate the performance of the the Pall Nucleic acid binding (NAB) Nanosep centrifugal device, which contains a dual layer silica-based quartz glass fiber media, for nucleic acid isolation and compare it with two other commercially available spin devices. All centrifugal devices chosen were silica-based spin devices and were
utilized for lysis-bind-wash-elute method of nucleic acid purification. One of the devices is not part of a kit and does not have its own buffers (herein will be referred to as ““AA”), whereas the other centrifugal device is from a total RNA isolation kit (herein will be referred to as “BB”) having its own buffers and manufacturer’s recommended protocol. Total RNA extraction buffers (herein will be referred to as Brand “X”) from a leading kit manufacturer were used to test performance of “AA” spin devices and NAB Nanosep spin device. The results indicate that the RNA isolated from E. coli TOP10 strain using Pall NAB Nanosep centrifugal device is either superior or equivalent to the commercially available brands AA and BB.