New England Biolabs (NEB), Ipswich, Mass, has released the NEBNext single-cell/low input RNA library prep kit for Illumina. The new kit incorporates an optimized template switching protocol—as well as NEB’s Ultra II FS enzymatic DNA fragmentation technology—to produce full-length transcript, sequence-ready libraries from single cells and ultralow input RNA.
“Studies based on population analyses using thousands of cells disguise the potentially significant biological variations among subpopulations of cells and in individual cells, and the benefits of sequencing the transcriptomes of these smaller samples is clear,” says Fiona Stewart, NEBNext portfolio manager at NEB. “However, there is a need for more sensitive, reliable, and affordable products for library preparation for single-cell and ultralow input RNA samples, in order to increase the breadth of transcripts detected, and the reliability of that transcript detection, as well as enabling distinction between sample variation and experimental variation.
“To address this growing need, we’ve incorporated a unique protocol and suite of reagents in our kit to reliably handle cultured or primary cells, with the flexibility also to accommodate as little as 2 pg, but up to 200 ng, of total RNA,” adds Stewart. “The inclusion of NEB’s Ultra II FS technology further streamlines the workflow and minimizes sample loss to enable robust full-length transcript, sequence-ready library generation.”
The NEBNext single-cell/low input RNA library prep kit meets the needs of operations at any scale and with a range of sample types, according to reports by early access users.
“We recently developed and implemented a novel single-cell transcriptomics pipeline to support our single-cell research initiatives, including the Human Cell Atlas project,” explains Peter Ellis, PhD, senior staff scientist at the Wellcome Sanger Institute. “Fully deployed, this high-throughput pipeline will be capable of generating single-cell RNA-seq data from approximately 30,000 samples per month. The scale of this operation is made possible by combining the streamlined workflows of the NEBNext single-cell/low input kit with our expertise in high-throughput liquid handling and sample processing.”
“The implementation of this new single-cell RNA-seq pipeline is an important addition to our core scientific operations and will enable our scientists to resolve at scale the transcriptional variation within a wide variety of single cells,” adds Sarah Teichmann, PhD, head of cellular genetics at Wellcome Sanger Institute.
The NEBNext single-cell/low input RNA library prep kit’s workflow includes new reagents, combined steps, and minimized cleanup steps, making it fast and easy to use, with less than 30 minutes hands-on time. Downstream of complementary DNA (cDNA) synthesis, sequencing library construction incorporates the Ultra II FS technology, allowing users to perform enzymatic cDNA fragmentation, end repair, and dA-tailing reactions in a single enzyme mix with a single protocol. A module with reagents only for cDNA synthesis
To learn more, visit New England Biolabs.