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REALSEQAC Sigma-Aldrich

RealSeq®-AC

NGS library preparation kits for miRNAs and small RNAs, For total RNA samples from tissue or cells

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Description

General description

RealSeq® is a novel method for preparing small-RNA sequencing libraries that reduces incorporation bias in Next-Generation-Sequencing (NGS). 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. This technology solves the problem of commonly used sequencing library preparations that lead to underdetection of many miRNAs.

RealSeq®-AC is a small RNA library preparation kit compatible with Illumina Next Generation Sequencing (NGS) technology. RealSeq®-AC is optimized for RNA input between 1ng and 1000ng of total RNA from tissue or cells.

Application

This kit is for small RNA library preparation for NGS. RNA-Seq for small RNA and miRNA; NGS library generation for small RNA and miRNA; RealSeq®-AC is highly efficient, detecting more miRNAs in total RNA samples.

Application Note
RealSeq® accurately quantifies small RNA sequenced on the Illumina Platform. RealSeq®-AC uses ligation of a single adapter producing small RNA libraries with more miRNAs compared to library prep kits using two adapters.

Features and Benefits

• Significantly reduces sequencing bias in small-RNA sequencing
• Accurately quantifies biologically relevant small RNAs
• Allows for more efficient discovery of novel small RNAs

Components

This kit is for 12 libraries. This product is comprised of the following:
RNA buffer
RealSeq Adapter
RNAse inhibitor
Ligation buffer
Ligase
Blocking Agent
Blocking Enzyme
Blocking Buffer
RealSeq Enzyme
RealSeq Buffer
Dimer Removal Agent
RT Primer
dNTPs
RT buffer
RT Enzyme
PCR Buffer
PCR Polymerase
Rnase Free Water
RealSeq Beads
Reverse Index Primers*
Forward Primer
miRNA Control
Adapter Dilution Buffer

* Index Primers 1-12 are included in REALSEQAC-1KT (500-00012)

See experimental protocol for more information.

Storage and Stability

The three components should be stored at the specified Temperature upon arrival of the Kit.

• RealSeq®-AC (BOX 1) at −20 °C
• RealSeq®-AC (BOX 2) at −20 °C
• RealSeq®-AC (Tube with Beads 3) at 2-8 °C

Legal Information

RealSeq is a registered trademark of SomaGenics, Inc.

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.

Documents

Certificate of Analysis (COA)

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Protocols & Articles

Articles

Small RNA and miRNA (microRNA) Sequencing

The field of next-generation sequencing (NGS) has observed change at a rapid pace, with exponential gains in both data generation and scientific discoveries. The power of NGS lends itself to the exam...
Keywords: Amplification, Cancer, Diseases, Gene expression, Ligands, Microarray Analysis, Peptide synthesis, Polymerase chain reaction, Purification, Sequencing, Transcription

Protocols

RealSeq®-AC miRNA Library Kit for Illumina® Sequencing Protocol

Overview RealSeq®-AC Kit Contents Warnings and Recommendations User-supplied Reagents, Consumables and Lab Equipment (not included) Input Requirements Experimental Protocol Appendix A: Thermocycler P...
Keywords: Amplification, Molecular biology, Peptide synthesis, Polymerase chain reaction, Purification, Separation, Sequencing, Size-exclusion chromatography, Transcription

Related Content

Webinar: Decreasing miRNA sequencing bias using a single adapter and circularization approach

Presenter: Sergio Barberan-Soler, Ph.D., Director of Sequencing Technologies, SomaGenics, Inc.
Keywords: Peptide synthesis, Reductions, Sequencing

Peer-Reviewed Papers
15

References

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