MilliporeSigma
All Photos(1)

REALSEQAC

Sigma-Aldrich

RealSeq®-AC

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

shipped in

wet ice

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

Application

RealSeq®-AC has been used for the construction of sRNA libraries from Rosa chinensis, Aquilegia coerulea, Triticum aestivum, and Hordeum vulgare.

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.

Pictograms

Environment

Hazard Statements

Precautionary Statements

Hazard Classifications

Aquatic Chronic 2

Storage Class Code

10 - Combustible liquids

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

Enter Lot Number to search for Certificate of Analysis (COA).

Certificate of Origin

Enter Lot Number to search for Certificate of Origin (COO).

Articles

Small RNA and miRNA (microRNA) Sequencing

A review on reducing miRNA and other small RNAs sequencing bias with RealSeq®-AC RNA library preparation on the Illumina® RNA sequencing platform.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Contact Technical Service