New England Biolabs NEBNext Low-bias Small RNA Library Prep Kit

New England Biolabs (NEB) has launched the NEBNext Low-bias Small RNA Library Prep Kit to minimise biased representation of small RNA species in sequencing data. The kit’s next-generation small RNA preparation method is designed to be faster and less biased, with a broader input range than other commercially available kits.

Researchers have only recently realised that small RNA species — including rare and non-coding RNAs — have an outsized influence on health and disease. However, sequencing bias in pre-existing library preparation methods has made it hard for researchers to understand the true diversity and abundance of small RNAs in their samples. The prep kit was designed to generate RNA libraries with a more accurate representation of small RNA species.

Protocol enhancements include the addition of a novel splint adaptor that increases the diversity of interactions, facilitating ligation and increasing sensitivity, with a streamlined, simplified protocol. As a result, researchers can now analyse all RNA species present in biologically relevant samples. Additional improvements include high speed (~3.5 h), long shelf life (18 months) and wide input range. Standard and 2´-O-methylated samples can also be processed using the same protocol, with multiplexing enabled through up to 480 compatible UDI primer pairs (available separately).

The kit’s novel approach to adaptor ligation has culminated from over a decade of research by NEB scientists. The first study, published in 2012, recognised that RNA sequencing bias was significant and caused mostly by unpredictable differences in ligation efficiency for small RNAs. The second foundational study, published in 2020, presented a novel library preparation workflow that reduced bias and increased sensitivity in small RNA library prep. Several protocol enhancements have since improved the kit to outperform the originally published methods.