Thermo Scientific Next Generation Sequencing Library Preparation Products

• Fast and convenient NGS library preparation workflow• High-efficiency reagents• Reliable performance

Tune up your next generation library preparation workflow

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Thermo Scientific™ NGS library preparation, cleanup and size-selection products combine all necessary NGS library construction steps into one streamlined user-friendly workflow. Optimized for maximal performance and flexibility, Thermo Scientific™ reagents, in combination with Thermo Scientific™ MagJET™ and Thermo Scientific™ GeneJET™ NGS cleanup and size selection kits, allow for fast, effortless and reliable DNA library construction from minimal DNA sample input.

Highlights

• Fast and simple – streamlined nucleic acid purification, NGS library preparation, cleanup and size-selection workflows

• Versatile – flexibility to generate NGS fragment libraries from nanogram to microgram DNA sample input

• Efficient – reagents optimized for maximum performance, high quality and reliable results

Thermo Scientific NGS library preparation workflow

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GeneJET and MagJET nucleic acid purification kits for:

Cells and tissuesBloodPlants

Nucleic acid purification

NGS library preparation

Cleanup and size selection

To NGS workflow

• ClaSeek Library Preparation Kits

• MuSeek Library Preparation Kits

• MagJET NGS Cleanup and Size Selection Kit

• GeneJET NGS Cleanup Kit

A complete solution for NGS library preparation workflow

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ClaSeek Kits MuSeek KitsSample input 5 ng-1 µg physically sheared DNA 100 ng of intact genomic DNA, cDNA

Method Ligation-based Transposon-based

PCR-free protocol available Yes No

Physical sample fragmentation required

YesNo

(transposon-mediated)

Protocol~ 60 min PCR-free protocol

~120 min amplification protocol ~ 80 min total time

Multiplexing compatible Yes Yes

NGS platform compatibilityIllumina™ MiSeq™, HiSeq™,

Ion Torrent™ PGM™, Ion Proton™

Illumina™ MiSeq™, HiSeq™, Ion Torrent™ PGM™, Ion Proton™

NGS library preparation in tune with your needs

The Thermo Scientific™ product portfolio provides two distinct NGS library construction technologies, each created to meet researchers' demanding needs. The outstanding versatility of the Thermo Scientific™ ClaSeek™ kits and ease of use of the Thermo Scientific™ MuSeek™ kits enable scientists to achieve excellent results with minimal effort in a short time.

ClaSeek KitsFlexible – DNA library construction from nanogram to microgram amounts

PCR-free – amplification-free protocols for unbiased library construction

Fast and convenient – significantly shorter library preparation workflow, less hands-on time

MuSeek KitsEasy-to-use – no physical shearing, end-repair or ligation steps

Fast – library generation in as few as 80 minutes

Universal – one protocol for generating libraries with different length fragments

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ClaSeek Kits – outstanding flexibility

Thermo Scientific ClaSeek Library Preparation Kits for Illumina™ and Ion Torrent™ platforms are designed for fast, convenient and accurate DNA library construction from DNA sample input as low as 5 ng. Amplification-free protocols eliminate PCR-related bias, resulting in uniform data coverage and reliable sequencing results.

Features

• Flexible – DNA library construction from 5 ng to 1 µg input

• PCR-free – significantly minimized bias and improved data coverage with amplification-free protocol

• Fast and convenient – library construction in only 60 minutes with minimal hands-on time

High data coverageSequencing data obtained from a DNA fragment library prepared using the ClaSeek PCR-free protocol demonstrates better sequence coverage in AT- and GC-rich DNA regions as compared to other suppliers. The Ion Torrent™ compatible DNA library was prepared from a mix of different genomic DNAs (100 ng) with varying GC content (T. thermophilus – GC ~ 69%; S. aureus –GC ~ 33%; E. coli – GC ~ 51%).

Ion Torrent™

and Illumina™ sequencing compatible

Barcoding compatible

DNA input as low as

5 ng

Fast and convenient workflowClaSeek Library Preparation Kits in combination with MagJET NGS Cleanup and Size Selection Kit enable fast and convenient DNA library preparation in as fast as one hour. The use of NGS-optimized reagent master mixes eliminates unnecessary pipetting steps and reduces hands-on time. Optional amplification protocols allow for DNA library preparation from sample input as low as 5 ng.

Libraries of different DNA fragment lengthThe ClaSeek workflow in combination with the MagJET NGS Cleanup and Size Selection Kit allows for preparation of DNA libraries comprising fragments of required fragment length. The image shows three Illumina™ sequencing compatible DNA libraries (input – 50 ng fragmented E. coli DNA) prepared for 50-, 300-, and 500-base sequencing runs on the Illumina™ MiSeq™ instrument. DNA fragment size distribution was analyzed with an Agilent™ Bioanalyzer™ 2100 using the High Sensitivity DNA kit.

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other suppliersThermo Scientific

MagJET cleanup

End conversion and adapter addition

MagJET size selection

PCR-free protocol ~ 60 minutes

Optional amplification protocol ~ 110 minutes

PCR amplification(Optional)

• Learn more: thermoscientific.com/claseek

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MuSeek Kits – speed and convenience

The Thermo Scientific MuSeek Library Preparation Kits utilize an innovative transposon-based technology that allows for enzymatic fragmentation and simultaneous tagging of the target DNA, eliminating the need for physical shearing, DNA end repair and adaptor ligation steps. This enables library construction in as few as 80 minutes with minimal hands-on time.

100 ng DNA

sample input

Features

• Easy-to-use – no physical shearing, end repair or ligation steps

• Fast – generates an NGS fragment library in as few as 80 minutes

• Universal – same protocol for generating different length fragment libraries

• Learn more: thermoscientific.com/museek

One protocol for different fragment length librariesThe MuSeek workflow in combination with E-Gel size selection tools (Life Technologies Inc.) enables DNA library preparation of different fragment length. The image shows a relative size distribution of an E. coli genomic DNA fragment library generated following the MuSeek protocol. Fragment libraries of target length were size selected from the same DNA fragment library. Fragment size distribution was analyzed with an Agilent™ Bioanalyzer™ 2100 using the High Sensitivity DNA Kit.

MuSeek worfklow for Ion Torrent compatible NGS library preparation.

35 100 150 200 300 400 500 600 1000 2000 10380 [bp]

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DNA fragment librarybefore size selection

DNA fragment librariesafter size selection

Bead purificationAdaptor addition PCRBead purification

5 min 5 min 5 min 5 min 20 min

10 min 20 min 30 min 20 min 80 min

Fragmentation Total time

Procedure time

Hands-on time

Size selection and continuation to Ion PGM workflow

Barcoding compatible

Ion Torrent™ and Illumina™

compatible

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Reliable NGS library cleanup and size selection

Thermo Scientific MagJET and GeneJET kits for NGS applications are uniquely optimized for robust and efficient DNA fragment library cleanup and size selection from a variety of reaction mixtures used in NGS workflows.

Features

• Robust – reliable cleanup and size selection from different reaction mixtures and buffers

• NGS-grade purity – complete removal of NGS adapters, primers, nucleotides, enzymes and other reaction components inhibiting the NGS workflow

Compatible with reaction mixtures and

buffers forDNA end conversion,

ligation, PCR, fragmentation and

elution

High yields and purityThe GeneJET NGS Cleanup Kit delivers higher yields of adapter-free DNA fragment library as compared to other suppliers. Fragmented DNA (1 µg) spiked with 50 bp length adapters (7 µg) was purified using Thermo Scientific and other supplier protocols. DNA yield was determined using the Agilent™ Bioanalyzer™ 2100. 1 – Fragmented DNA (control); 2 – Thermo Scientific Cleanup protocol; 3 – Thermo Scientific Adapter Removal protocol; 4-5 – other suppliers' kits.

Compatibility with different reaction mixtures and buffersMagJET and GeneJET kits demonstrate protocol compatibility with different reaction mixtures and buffers used in NGS workflows. Fragmented E. coli genomic DNA (1 µg) samples, each spiked into different reaction mixtures (a-h), were size selected using the MagJET protocol. Isolated fragment libraries were analyzed using the Agilent™ Bioanalyzer™ 2100. Expected median fragment sizes are marked with a dotted line. a – End conversion reaction mixture (Ion Torrent compatible); b – PCR reaction mixture (1); c – PCR reaction mixture (2); d – Ligation mixture; e – End conversion reaction mixture (Illumina compatible); f – MuSeek fragmentation reaction buffer; g – Elution buffer; h – Nebulization buffer; i – Control (input DNA).

MagJET NGS Cleanup and Size Selection Kit:• Size-selection of required DNA

fragment range• High-throughput compatible

magnetic bead technology• Automation on Thermo Scientific™

KingFisher™ instruments

GeneJET NGS Cleanup Kit:• Silica membrane-based nucleic

acid purification technology• Micro-column format• Fast 5-minute cleanup protocol

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Thermo Scientific Supplier A Supplier B

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Higher data output in RNA-seq applications

The Thermo Scientific NGS product portfolio provides all essential components to build a complete RNA-seq workflow from RNA isolation to NGS library construction. A combination of Thermo Scientific MagJET mRNA enrichment and Thermo Scientific™ Maxima™ H Minus double-stranded cDNA synthesis technologies ensure maximum output of meaningful sequencing data.

Higher mRNA read output in NGSA ds cDNA fragment library prepared from HeLa mRNA isolated using the Thermo Scientific MagJET mRNA Enrichment Kit demonstrates higher mRNA read output in RNA-seq applications than with other suppliers' kits. Four ds cDNA libraries were prepared using the ClaSeek Library Preparation Kit, Illumina compatible, and other suppliers' kits for mRNA enrichment from total RNA samples. ds cDNA synthesis was performed using the Maxima H Minus Double-Stranded cDNA Synthesis Kit.

MagJET mRNA Enrichment Kit• Up to 10-fold mRNA enrichment• Full-length high-integrity mRNA isolation• High yields from broad (5-100 µg)

RNA input range• Compatible with high-throughput

automation

Maxima H Minus Double-Stranded cDNA Synthesis Kit• Efficient synthesis of full-length,

double-stranded cDNA• Procedure completed in less

than two hours• Premixed components• Includes primers, controls and

residual RNA removal reagents

Thermo Scientific RNA-seq workflow

100%

80%

60%

40%

20%

0%Thermo

ScientificTotal RNA

Supplier A

mRNA rRNA/tRNA

Supplier B

RNA

read

s, %

To NGS workflow

Library preparation

MuSeek and ClaSeek Library Preparation Kits

RNA isolation mRNA enrichment

ds cDNA synthesis

GeneJET and MagJET Total RNA Purification Kits

MagJET mRNA Enrichment Kit

Maxima H Minus Double-Stranded cDNA Synthesis Kit

• Learn more: thermoscientific.com/ngs

Ordering details

Product Size Cat. No.

MuSeek Library Preparation Kit for Ion Torrent 10 rxns K1331

MuSeek Library Preparation Kit, IlluminaTM compatible 12 rxns K1361

MuSeek Barcode Set 1, Ion Torrent compatible 80 rxns K1541

MuSeek Index Set 1, Illumina compatible 96 preps K1551

ClaSeek Library Preparation Kit, Illumina compatible12 rxns K1341

48 rxns K1342

ClaSeek Library Preparation Kit, Ion Torrent compatible10 rxns K1351

50 rxns K1352

GeneJET NGS Cleanup Kit50 preps K0851

250 preps K0852

MagJET NGS Cleanup and Size Selection Kit

1 × 96 preps K2821

4 × 96 preps K2822

10 × 96 preps K2823

10 preps K2828

MagJET Separation Rack

2 × 1.5 mL MR01

12 × 1.5 mL MR02

4 x 50 mL MR04

MagJET mRNA Enrichment Kit10 preps K2811

96 preps K2812

Maxima H Minus Double-Stranded cDNA Synthesis Kit 10 rxns K2561

© 2014 Thermo Fisher Scientific Inc. All rights reserved. Agilent and Bioanalyzer are trademarks of Agilent Technologies and its subsidiaries. E-Gel, Ion Proton, Ion Torrent and PGM are trademarks of Life technologies and its subsidiaries. Illumina, MiSeq, HiSeq are trademarks of Illumina and its subsidiaries. All other trademarks are the property of Thermo Fisher Scientific Inc. and its subsidiaries.

Europe

Customer Service cs.molbio.eu@thermofisher.com

Technical Support ts.molbio.eu@thermofisher.com

Tel 00800 222 00 888 Fax 00800 222 00 889

United States

Customer Service cs.molbio@thermofisher.com

Technical Support ts.molbio@thermofisher.com

Tel 800 235 9880 Fax 800 292 6088

Canada

Customer Service cs.molbio@thermofisher.com

Technical Support ts.molbio@thermofisher.com

Tel 800 340 9026 Fax 800 472 8322M

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