RealSeq Small RNA Library Preparation Kits
RealSeq’s industry leading small RNA NGS library technology allows broad detection with “best in class” low bias. Their wide range of sequencing services includes library preparation from extracted RNA or ready-to-sequence pooled libraries as well as customization for your bioinformatic needs.
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RealSeq® technology for small RNA sequencing library preparation
Accuracy through circularization
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.
The Company is the recipient of several NIH SBIR Phase I/II grants including fundng for its RealSeq NGS library construction platform.
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.
See our publication: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6120088/
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.
Targeted Approach
RealSeq-T is a novel technology that allows the targeted, direct quantification of microRNAs from biofluid samples. Data from plasma samples show that employing a targeted sequencing approach eliminates the need to perform RNA extraction and is significantly more reproducible compared to non-targeted approaches that detect microRNAs after standard RNA extraction.
Single Cell
RealSeq®-SC is an innovative and highly efficient technology to improve the detection accuracy of single-cell small RNA sequencing and minimize technical dropout rates. The technology expands single-cell transcriptomics to encompass small as well as large RNAs, is effective across multiple cell types, and provides an effective platform for single cell biomarker/liquid biopsy applications.
Selected Suppression Probes – SSP
Highly-expressed RNA species reduce read depth of less abundant species that researchers care about. RealSeq Biosciences has developed a targeted removal probe (Selective Suppression Probe- SSP™) approach to depleting over-abundant species from RNA samples, resulting in increased miRNA reads and greater sequencing depth of miRNAs.
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Ordering information
RealSeq®-AC
Complete Small RNA library preparation kit for Illumina for RNA samples from tissues or cells. Patented technology with a single-adapter and circularization to reduce detection bias. For total RNA samples from tissue or cells (1 ng to 1000 ng total RNA input). Size selection reagent and barcoded PCR primers are included.
RealSeq®-Biofluids
Complete Small RNA library preparation kit for Illumina for RNA samples from biofluids and other low concentration samples. Patented technology with a single-adapter and circularization to reduce detection bias. For RNA samples extracted from biofluid samples and other sample types with low RNA concentration. Size selection reagent and barcoded PCR primers are included.
RealSeq® Dual Index 12RXN
Complete Small RNA library preparation kit for Illumina for RNA samples from tissues or cells. Patented technology with a single-adapter and circularization added capability of using dual barcoding. Allows multiplexing of up to 384 samples on the same sequencing run.
RealSeq® Size Select Beads
Size selection beads to purify libraries after PCR. Already included with all our kits, order this when extra purifications are required.
RealSeq® Index Primers
Barcoded PCR primers sets, with 12 barcodes per set. Order when additional barcodes are required.
miND® spike-in
miND® Spike-Ins are a novel quality control parameter and normalizer that consists of seven oligonucleotides, each characterized by a unique core sequence flanked by 4 randomized nucleotides. miND® Spike-Ins are provided in a specific ratio to cover the broad concentration range of endogenous small RNAs.
Selective Suppression Probes – SSP™
Deplete unwanted or over-abundant RNA fragments from your samples. SSPs can reduce unwanted rRNA reads to about 0.1% of total reads while increasing small RNA/miRNA reads by 50%.
Human Plasma Panels and custom probes are available. Please inquire: support@realseqbiosciences.com
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