RealSeq®-Dual miRNA Library Kit for Illumina® sequencing

Complete Small RNA library preparation kit using dual barcoding

Product Description

• RealSeq-Biofluids Kit allows preparation of gel-free sequencing libraries with an RNA input obtained from only 5 µl of plasma (optimal 200 µl)
• RealSeq-Biofluids delivers the highest percentage of usable reads (>15 nt and that align to either miRBase or genome) of the 3 kits tested
• Highly accurate profiling allows the identification of a larger set of cf- miRNAs
• Detection bias inherent in the two-adapter platforms reduces the number of miRNAs identified
• Detection bias also results in the overrepresentation of a few miRNAs that consume the majority of sequencing reads
• Allows multiplexing of up to 384 samples on the same sequencing run.

Workflow

Supporting Data

Competitor Performance Comparison

Fig.1. The same plasma sample was used to prepare sequencing libraries with three different kits. Graph shows sequencing metrics of libraries prepared with each kit. Short reads correspond to reads <15 nt after adapter trimming; reads passing this filter are then align to a reference file with all human miRNAs (miRBase 21), reads that do not align to miRNAs are then aligned to the human genome (hg19).

Detection of plasma miRNAs with different library preparation kits

Fig.2. Comparison of miRNAs read frequencies of three different library preparation kits (gel-free protocols) from 200 ul plasma samples. Reads were subsampled to 10 million reads per kit and aligned to a reference that includes all human miRNAs listed in miRBase 21 (Miltenyi Biotec). The number of miRNAs detected at different coverage for each library preparation kit is shown. Especially low copy-number miRNAs are significantly more sequenced when the RealSeq® - biofluids kit was used for library preparation.