Tel: +44 (0)1223 316 855
life science research products, biological research products, biotechnology
close ...


Supplier:
Product types:

RealSeq-Biofluids

Find Your RNAi Quickly with Our RNAi Search
Powerful RNA Extraction From FFPE Samples
Achieve High Yield Mammalian Protein Production In Suspension HEK 293 & CHO Derived Cells

RealSeq®-AC small RNA library kit for Illumina® sequencing

RealSeq®-AC small RNA library kit for Illumina® sequencing

RealSeq®-AC small RNA library kit offers a novel, proprietary method for preparation of small RNA sequencing libraries that minimises bias related to intramolecular RNA folding and adapter dimers. Offering an efficient, gel-free method which detects more miRNAs in total RNA than other miRNA library preparation kits and optimised for inputs between 1 ng - 100 ng total RNA, the RealSeq®-AC small RNA library kit for Illumina® sequencing is the ideal solution for small RNA library preparation.

Download the ABRF 2018 poster
Decreasing miRNA sequencing bias using single adaptern and circularisation approach

Benefits of using this kit
• Eliminates bias-induced over/under represented small RNAs
• Efficient and gel-free workflow
• Optimised for inputs between 1 ng - 100 ng total RNA
• Detects more miRNAs in total RNA than other miRNA library preparation kits
• Lower sequencing costs due to the reduced number of irrelevant sequencing reads

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-AC reduces sequencing bias in known problematic miRNAs

Spectrum of bias assessed using synthetic miRNAs. A commonly used two-adapter protocol is compared with RealSeq-AC’s one-adapter protocol. Typically under or over represented miRNAs are assayed and deviation from the optimal "0" value are shown. The RealSeq-AC method significantly reduces the detection bias.


RealSeq-AC reuces incorporation bias compared to other small RNA library preparation kits


Bias Measurements for various small-RNA library preparation kits. All sequencing libraries were prepared in triplicate using 1 pmole of miRXPlore Universal Reference pool (Miltenyi Biotec), which consists of equimolar concentrations of 963 synthetic mature miRNAs. Purified libraries were sequenced on the Illumina MiSeq platform. Trimmed sequencing reads were aligned to a custom miRNA reference (Bowtie2, Langmead et al. 2012). Reads mapping to miRNAs were counted and fold-deviations from the equimolar input were calculated and plotted as log2 values. Measurements of miRNA levels falling within a factor of two of the expected values (between vertical lines) are considered unbiased according to Fuchs et al (2015). The method of adapter attachment to the miRNAs is noted under the graphs.

Material available for download
RealSeq®-ACmiRNA 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

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by SomaGenics, Inc. No right under any other patent claims to methods, apparatus or reagents is conveyed expressly, by implication, or by estoppel. This product is intended for research use only and may not be transferred to third parties, resold, modified for resale or used to manufacture commercial products without prior written approval of SomaGenics, Inc. For more information about commercial rights, please contact SomaGenics Business Development at bd@somagenics.com.

Products

Note: availability depends on country - see product detail page.

View product detail
Applications: preparation for Next-generation sequencing of small RNAs
View product detail
Applications: preparation for Next-generation sequencing of small RNAs
View product detail
Applications: preparation for Next-generation sequencing of small RNAs
Chat To Our Somagenics Specialist Emma