The RealSeq®-AC kit from Somagenics offers a proprietary method for the preparation of small RNA sequencing libraries for Illumina® sequencers, that minimises bias related to intramolecular RNA folding and adapter dimers. By using an efficient, gel-free approach, this technology detects more miRNAs in total RNA than other miRNA library preparation kits and is optimised for inputs between 1 ng - 100 ng total RNA.
Benefits of using the RealSeq-AC small RNA library kit
• Eliminates bias-induced over/under-represented small RNAs
• Accurately quantifies biologically relevant small RNAs
• Optimised for inputs between 1 ng - 100 ng total RNA
Download the ABRF 2018 poster:
Decreasing miRNA sequencing bias using single adaptern and circularisation approach
RealSeq technology schematic
RealSeq-AC technology detects more miRNAs in total RNA* than other miRNA library preparation kit
|Kit I||Kit N||Kit B||Kit T||RealSeq-AC|
|miRNAs with > 5 reads||404||452||412||324||500|
|miRNAs with > 10 reads||328||365||352||239||385|
RealSeq-AC detects more miRNAs with over five or ten reads per million, respectively, from total RNA samples compared to other kits for miRNA sequencing library preparation.
RealSeq accurately quantifies small RNAs
Bias measurements comparing small RNA library preparation kits. 1 pmol of miRXPlore Universal Reference pool (Miltenyi Biotec) was used to compare incorporation bias in six different library preparation kits. Purified libraries were sequenced on the Illumina MiSeq platform. Trimmed sequencing reads were aligned to a custom miRNA reference. Reads mapping to miRNAs were counted and fold-deviations from the equimolar input were calculated and plotted as log2 values. Measurements of miRNA levels within a factor of two of the expected values (between vertical lines) are considered unbiased. The method of adapter attachment to the miRNAs is noted in the legend. Libraries prepared using RealSeq®-AC have substantially fewer miRNAs with bias than those prepared using any of five other commercially available library preparation kits.
Material available for download
RealSeq-AC miRNA Library Kit for Illumina® sequencing
RealSeq-AC Low Quality RNA application note
RealSeq-AC Size Selection with Novex Gels application note
RealSeq-AC Low Input application note
Decreasing miRNA sequencing bias using a single adapter and circularisation approach
Assessment of cell-free nucleic acid isolation efficiencies for liquid biopsy research
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