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Small RNA-seq

Small RNA Library Kits for Illumina Sequencing

Identify and quantitatively analyze the entire population of small RNAs

Find the right system for your Small RNA NGS Library Preparation project

  • The NEXTFLEX™ Small RNA Sequencing Kit v4 from Revvity is a gel free, automation ready platform for diverse sample types, including challenging biofluids and exosomes, with strong bias and dimer reduction. The kit utilizes an adapter-ligation strategy.
  • RealSeq® Kits from RealSeq Biosciences apply a patented single adapter circularization technique to reduce detection bias. Kits are offered for tissues/cells (RealSeq® AC), delivering TruSeq compatible analysis and Biofluids (RealSeq® Biofluids), which is a plasma/serum focused cf miRNA system for extremely low concentration biofluid RNA.  
  • All-inclusive Transcript/RNA Fragment Profiling: Detect small RNAs, diverse RNA fragments (including lncRNA, mRNA and tRF/tDR), and select modifications in a single reaction with the RiboMarker RNA Fragmentomics Library Preparation Kit from RealSeq Biosciences. 

NEXTFLEX NGS Library Preparation

The NEXTFLEX Small RNA Seq Kit v4 is a small RNA library prep solution that delivers low bias, low dimer libraries from as little as 1 ng total RNA across tissues, biofluids and exosomes, with strong miRNA discovery performance and 384 plex UDI multiplexing for Illumina and Element sequencing. 

The kit is automation-ready on Sciclone™ G3 NGSx, Sciclone G3 NGSx iQ™, and Zephyr™ G3 NGS workstations for walk-away scalability.

All of Revvity’s NGS adapters incorporate unique dual indexes (UDI) and are compatible with Illumina, Watchmaker, Twist, and any other TruSeq style library prep kit. We also offer adapters with both unique molecular adapters (UMI) and UDIs. Incorporating full-length UDI and UMI sequences via ligation (instead of PCR) reduces the risk of introducing errors.

NEXTFLEX® Small RNA Sample Preparation Flow Chart

Nex Smallrna Seq Kit How It Works


RealSeq® small RNA/ miRNA Library Kits

RealSeq Biosciences’ core expertise includes innovative proprietary technologies for bias-free small RNA/miRNA NGS library construction, targeted NGS tools, and cf-RNA analysis (liquid biopsy) that form the basis of the Company’s life science research programs and product development. Two Kits for different sample types are available:

  • RealSeq Biofluids is a plasma/serum optimized, gel free small RNA library kit for Illumina that uses single adapter circularization, HY4 and dimer removal to deliver low bias cf miRNA and ncRNA profiling from as little as 50–200 µl of biofluid.
  • RealSeq AC is a TruSeq compatible small RNA library kit for Illumina that supports 1 µg–1 ng high quality total RNA with defined adapter/PCR settings, integrated miRNA controls and indexed primers for up to 48 sample multiplexing.

The RealSeq Technology solves the problem of commonly used sequencing library preparations that lead to underdetection of many miRNAs, some by as much as 10,000-fold. By using a novel single adapter and circularization approach, RealSeq® greatly reduces library preparation bias.

For further improved sequencing results RealSeq has developed  a targeted, customizable removal probe approach (Selective Suppression Probes- SSPs) that depletes uninteresting, over-abundant RNA fragments from your samples.

RealSeq® Technology

Realseq Ac Scheme

Sergio Barberán-Soler*, Jenny M. Vo, Ryan E. Hogans, Anne Dallas, Brian H. Johnston and Sergei A. Kazakov* (2018) Decreasing miRNA sequencing bias using a single adapter and circularization approach. Genome Biology, 19:105.