New England Biolabs (NEB) has introduced the NEBNext Low-bias Small RNA Library Prep Kit, a new solution designed to reduce bias in small RNA sequencing data. This kit offers faster processing times, a wider input range, and improved shelf life compared to other kits on the market.

Small RNA species, including rare and non-coding RNAs, play significant roles in health and disease. However, previous library preparation methods often introduced sequencing bias, making it difficult for researchers to accurately profile the diversity and abundance of these RNAs.

Betsy Young, PhD, senior product marketing manager for the NEBNext portfolio, said the kit was developed to provide a more precise representation of small RNA species. She explained that the kit’s novel splint adaptor enhances ligation sensitivity by increasing the diversity of interactions. The streamlined protocol enables researchers to analyze all RNA species in biologically relevant samples more confidently.

The new kit completes the library preparation process in approximately 3.5 hours and has an 18-month shelf life. It supports both standard and 2´-O-methylated RNA samples using a single protocol. Additionally, the kit allows multiplexing with up to 480 unique dual index (UDI) primer pairs, which are available separately.

The development of this kit builds on over ten years of research by NEB scientists. Initial studies published in 2012 identified significant bias caused by differences in ligation efficiency during small RNA sequencing. A subsequent study in 2020 introduced a novel library preparation workflow that reduced bias and increased sensitivity. Since then, several enhancements have further improved the kit’s performance.

Greg Lohman, research group leader in NEB’s molecular enzymology division, highlighted the impact of sequencing bias on RNA research. He said resolving this bias will facilitate more diverse RNA studies and help enable future scientific discoveries.

For more information about the NEBNext Low-bias Small RNA Library Prep Kit or to request a trial, visit https://www.neb.com/en-us/products/e3420-nebnext-low-bias-small-rna-library-prep-kit.