New England Biolabs, Inc. (NEB) launched their new NEBNext® "Ultra" kits at the recent American Society for Human Genetics (ASHG) annual meeting. These kits provide streamlined, low-input methods to prepare DNA and RNA libraries for Illumina® next generation sequencing. The supplied protocols and reagents are designed to maximize useful data from a broad range of samples, including those available in limited amounts.
The NEBNext Ultra DNA and Ultra RNA Library Kits produce high-yield libraries from 5 ng to 1 ug of input DNA, or as little as 10 ng of input RNA. The input RNA can be total RNA, purified mRNA or rRNA-depleted RNA.
Early access user Cynthia Hendrickson, Ph.D., at the HudsonAlpha Institute for Biotechnology, reports that in initial experiments, NEB's new method "has allowed us to reduce our DNA inputs for exome enrichment from micrograms to a few hundred nanograms, while providing a simpler, faster protocol that minimizes the amount of hands-on time required."
Another early access user, Momchilo Vuyisich, Ph.D., at the Los Alamos National Laboratory, says, "After testing several NGS library prep kits, we have found that the NEBNext Ultra DNA kit for Illumina has the best overall utility for resequencing and assembly of bacterial genomes. The kit offers an unmatched combination of ease-of-use, low cost, robustness and low input DNA requirements. It consistently produces excellent sequencing data."
The new Ultra kits contain novel ligation reagents, as well as NEB's NEBNext NGS-optimized formulation of Q5® High-Fidelity DNA polymerase, which provides ultra high-fidelity amplification and minimized GC bias.
NEB's streamlined protocol for constructing DNA libraries with the Ultra kit requires only 15 minutes of hands-on-time, and is complete in 2.5 hours. The Ultra RNA workflow also incorporates a streamlined protocol; the workflow is complete in 4-5 hours, with only 30 minutes of hands-on time.