Small RNA-seq

Small RNA Library Kits for Illumina Sequencing

Next Generation Sequencing (NGS) is a powerful tool to identify and quantitatively analyze the entire population of small RNAs

RealSeq® small RNA/ miRNA Library Kits

Accuracy Through Circularization - Bias-free small RNA/miRNA NGS library construction

  • Elimination of incorporation bias
  • Detect all important biomarkers
  • Gel-free highly efficient protocol yielding superior quality sequencing libraries
  • Lower sequencing costs by reducing the number of irrelevant reads
  • Kits for low total RNA inputs from either cells/tissue or biofluids such as plasma/serum

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. 

RealSeq® is a novel method for preparing small-RNA sequencing libraries that nearly eliminates incorporation bias in Next-Generation-Sequencing (NSG). This 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.

Under-representation in sequencing libraries can obscure the presence of some RNAs, including potential biomarkers. Accurate quantification of microRNAs (miRNA) and other small RNAs is important for understanding their biology and for developing new biomarkers and therapeutic targets.

Most bias stems from sequence-dependent variability in the enzymatic ligation reactions that attach the two adapters to the 3’ and 5’ ends of the miRNAs /small RNAs during preparation of sequencing libraries. By using a novel single adapter and circularization, RealSeq® greatly reduces library preparation bias.

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.

More Small RNA Library Preparation Kits

developed  the RapidSeq Small RNA Sample Prep Kits for preparation of NGS libraries for subsequent cluster generation, using purified small RNA or total RNA which contains small RNA fragments as input. The protocol includes steps for adapter ligation, reverse transcription, PCR amplification, and size selection by gel purification to generate a library product compatible with the Illumina NGS platform. 4 kits with different sets of 12 aligners, respectively, are available.

The Small RNA Library Prep Kit for Illumina developed by Norgen Biotek is optimized for the generation of small RNA-seq libraries from bodily fluids. It comprises all the reagents and components required to generate small RNA libraries to be used for next generation sequencing on an Illumina platform. A purification module is also provided for rapid purification of nucleic acid products generated at various steps of the workflow. The purification module utilizes Norgen’s patented resin technology which enhances recovery of desired library intermediates or final products. The library prep workflow could be used for different forms of input including purified total RNA or enriched small RNA, as well as RNA from low content inputs such as plasma, serum and urine.