Good protocols are the currency of any lab. If your research involves working with small RNAs, be sure to check out this carefully documented protocol published in Extracellular RNA, part of Springer Nature’s Methods in Molecular Biology book series.
The protocol can be found in the chapter “Preparation of Small RNA NGS Libraries from Biofluids.” Authors Alton Etheridge, Kai Wang, David Baxter, and David Galas present an NGS protocol designed for use with low-input biofluid samples containing extracellular RNAs. The protocol “has modifications designed to reduce the sequence-specific bias typically encountered with commercial small RNA library construction kits,” the authors report. The result is increased library diversity and more complete RNA profiles.
As the writers note, extracellular RNA has great promise for use as a biomarker or diagnostic, offering a non-invasive approach to everything from monitoring cancer to prenatal testing. The challenge for scientists, though, is that typical small RNA library prep kits require higher concentrations than can be gleaned from biofluid samples such as plasma or serum. This new protocol optimizes steps for samples that do not meet those concentration thresholds.
We’re pleased to see that PippinHT is featured heavily in the protocol for DNA sizing steps following PCR amplification. Indeed, the authors report that “use of an automated size selection instrument like the [Pippin Prep], BluePippin, or PippinHT can reduce variability introduced during gel excision.” We couldn’t agree more!