Introducing the NEBNext® Low-bias Small RNA Library Prep Kit
By Marlene L. Nielsen
Bias in small RNA library prep can seriously impact the accuracy of your data, masking rare RNA species. The NEBNext Low-bias Small RNA Library Prep Kit minimizes biased representation of small RNAs, more accurately reflecting the number and proportion of unique small RNAs present in your sample. NEB’s next-generation kit utilizes a streamlined workflow to capture small RNAs (<120 nt) with a 5’ phosphate and 3’ hydroxyl group, and can be used to identify microRNAs (miRNAs), transfer RNAs (tRNAs) and tRNA-derived fragments, small nucleolar RNAs (snoRNAs), piwi-interacting RNAs (piRNAs), and plant miRNAs. The kit is available in two versions: 24 rxn (NEB-E3420S) and 96 rxn (NEB-E3420L).
Key features
- Flexible input:
- 1,000–0.5 ng Total RNA
- 5–0.05 ng Enriched Small RNA
- One single protocol for both standard and 2′-O-methylated RNAs
- Multiplexing-ready: compatible with up to 480 UDIs using NEBNext LV Unique Dual Index Primers Set 1, 2, 2A, 4, 5 (NEB-E3400S, NEB-E3402S, NEB-E3390S, NEB-E3406S, NEB-E3408S).
- No gels required: two bead-based cleanups
- Fast workflow: complete prep in ~3.5 hours
Performance highlights
Lowest bias
Using a panel of 100 synthetic miRNAs (including 2′-O-methylated ends), libraries generated with the NEBNext kit showed:
- 90% of sequences within 2-fold of expected values
- Competitor kits showed only 19–30% within range
- Lower proportion of underrepresented sequences (15–24%) vs. 50–65% for other kits
Higher yields, fewer cycles
Libraries prepared from 0.5–1,000 ng human brain RNA yielded results equal to or better than those from other kits, while requiring 2–5 fewer PCR cycles. Fewer cycles reduce amplification bias and preserve complexity.
Want to learn more?
Download the Product Brochure and Manual or try a free sample: E3420
NEB offers a wide variety of Free of charge Samples covering a whole range from nucleic acid purification, PCR amplification to DNA assembly/cloning, as well as NGS library preparation. Check them out here
Want to know more? Reach out to Marlene!
