Small RNA Sequencing

Small RNA Sequencing

The Small RNA-Seq Library Prep Kit for Illumina consists of all the reagents and components required to generate small RNA libraries to be used for Next Generation Sequencing on an Illumina platform. All molecular reagents including adapters, primers, enzyme mixes, and buffers are provided. A column-based purification is included for rapid purification of nucleic acid products generated during the workflow.

Small RNA-Seq Library Prep Kit for Illumina

The Small RNA-Seq Library Prep Kit provides a protocol for generating small RNA libraries for Illumina sequencing, directly from total RNA or enriched small RNA. It enables discovery and profiling of all common types of small RNA’s (≤ 200nt), including miRNA, piRNA, siRNA, snoRNA, tsRNA, srRNA and U-RNA.

  • Gel-free user-friendly protocol
  • Wide input range from 50pg to 1µg
  • Optimized for low RNA-content samples such as plasma, serum and urine
  • Ready-to-sequence libraries in less than 5 hours
  • Multiplexing of up to 96 samples
Small RNA-Seq Library Prep Kit for Illumina
Exceptional miRNA discovery
figure 1: Total number of miRNAs detected across 4 different protocols. Dilution series (6pg, 60pg and 600pg) of purified plasma RNA used for library prep from different vendors were performed. The obtained libraries were sequenced at equal molarity at 1.

Exceptional miRNA discovery

The protocol allows for detection of a higher number of microRNA’s compared to other workflows. The difference is especially considerable for lower RNA inputs.

Optimized for RNA from challenging Sources

The high sensitivity of the kit makes it the best choice for challenging, low content RNA sources, such as liquid biopsies (plasma, serum and urine), including exosomes. The protocol has also been tested on RNA from cells and tissues and can be used for inputs from 1ng-1µg of cellular total RNA or 50pg-1µg of enriched small RNA, including liquid biopsies.

High Reproducibility

The protocol exhibits exceptional reproducibility and correlation across samples with different concentrations.

figure 2: a: Dilution series of purified plasma RNA in small RNA-seq shows high inter-replicate correlation reads pr Million (RPM) across concentrations tested (60pg and 600pg) b: Dilution series of purified plasma RNA in small RNA-seq shows high correlat

Gel-free User-friendly and Fast protocol

The Small RNA-Seq Library Prep Kit for Illumina offers a time saving protocol that can be completed within 5 hours and requires just 1h15’ of hands-on time. The final library does not need to go through gel purification. Quick and convenient magnetic bead-based purification can optionally be performed to remove adapter dimers for demanding RNA inputs.

Multiplexing of up to 96 samples

With the 96 i7 indices, standardly included in the kit, multiplexing of up to 96 samples is possible. This allows to pool more samples per sequencing lane and to perform cost-efficient experiments on the platforms of different scale.

Lexogen

Complete solution for Small RNA analysis

In order to get high quality input material for the Small RNA Library Prep, we recommend to extract the RNA with the SPLIT RNA extraction kit. This kit enables extracting NGS-grade total RNA or small and large RNA fractions from the same sample. SPLIT is highly suitable for enrichment of small RNA for use with the Small RNA-Seq Library Prep Kit.

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SPLIT RNA Extraction Kit

The SPLIT RNA Extraction Kit enables a fast and highly efficient extraction of RNA that is free of genomic DNA contamination. The RNA can be recovered as total RNA or split into two fractions, large RNA and small RNA, facilitating the analysis of e.g., mRNA and miRNA from the same sample. The RNA obtained is ideal for seamlessly preparing libraries for Next Generation Sequencing of small RNA’s.

Ordering information:

Cat. no.: 008.48 - Description: Split RNA extraction Kit - Reaction size: 48rxn

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Split RNA extraction kit
Lexogen

Gel Extraction Module

The Gel Extraction Module enables the size selection of Small RNA-Seq libraries in a convenient spin column format. It prevents unwanted sequences (e.g., linker-linker artifacts and longer library fragments) dominating the NGS results and helps focus the sequencing depth on the desired microRNA fraction.

The module includes a reagent box consisting of Loading Dye, NGS MW Ladder, and NGS Control Ladder, and a purification box with all necessary buffers and purification columns for clean-up.

Ordering information:

Cat. no.: 054.24 - Desciption: Gel Extraction Module - Reaction size: 24 rxn

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Purification Module with Magnetic beads 

The Purification Module with Magnetic Beads contains all the necessary reagents to carry out additional purifications.

Ordering information:

Cat.no.: 022.96 - Description: Purification Module with Magnetic beads - Reaction size: 200 rxn

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Contact information

+31 30 6880771
support@isogen-lifescience.com

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