NEXTflex™ Pre-Capture Combo DNA Library Prep Kit (NimbleGen SeqCap EZ Library Compatible)

Genomic DNA and exome library preparation for NimbleGen SeqCap EZ library construction and Illumina sequencing

NEXTflex™ Pre-Capture Combo Kit (48 barcodes)

Catalog# Product Name Quantity US List Price
5140-51 NEXTflex™ Pre-Capture Combo Kit (6 barcodes) 48 rxns $2,170 Buy Now
5140-52 NEXTflex™ Pre-Capture Combo Kit (12 barcodes) 96 rxns $4,229 Buy Now
5140-53 NEXTflex™ Pre-Capture Combo Kit (24 barcodes) 192 rxns $8,235 Buy Now
5140-56 NEXTflex™ Pre-Capture Combo Kit (48 barcodes) 384 rxns $14,752 Buy Now
5140-54 NEXTflex™ Pre-Capture Combo Kit (96 barcodes) 768 library prep rxns/192 target capture rxns $28,138 Buy Now

NEXTflex Pre-Capture Combo Kit (Roche NimbleGen SeqCap EZ Compatible)
  • Complete kit – Contains library prep, barcodes and barcode blockers compatible with NimbleGen’s SeqCap EZ Library target enrichment including exome sequencing
  • Increased multiplexing compatibility – 96 barcodes and barcode blockers available
  • Simplifies and reduces the cost of target capture
  • Optimized - Offers larger number of on-target reads
  • Completely gel-free library prep protocol available
  • Simplified workflow with master mixes reduces hands on time
  • Bead-based cleanup protocol
  • Compatible with Illumina sequencing platforms

The NEXTflex™ Pre-Capture Combo Kit (NimbleGen SeqCap EZ Library Compatible) is a complete Illumina-compatible library prep solution designed for DNA library preparation, multiplexing, and barcode blocking, upstream of Roche NimbleGen’s SeqCap EZ Library target enrichment. This kit facilitates deep multiplexing for both exome target capture and DNA target capture.

The NEXTflex PreCapture Combo Kit (NimbleGen SeqCap EZ Library Compatible) is designed to prepare single, paired-end, and multiplexed genomic DNA libraries for DNA and exome target capture and Illumina compatible sequencing. This kit features Enhanced Adapter Ligation Technology, resulting in library preps with a larger number of unique sequencing reads. With these improvements to the ligation enzymatic mix, the user will now have the ability to perform ligations with longer adapters and higher binding efficiencies. The NEXTflex Pre-Capture Combo Kit simplifies workflow by using master mixed reagents and magnetic bead based cleanup, reducing pipetting and eliminating time-consuming steps in library preparation. 


Automation Compatibility

The NEXTflex Pre-Capture Combo Kit is compatible with Beckman Biomek® FX and Biomek® FXP Laboratory Automation Workstations. A program is currently being validated for use with the Sciclone NGS, Sciclone NGSx and Sciclone G3  platforms. For more information contact NextGen@biooscientfic.com.


Increased Multiplexing Capabilities

The NEXTflex™ DNA Barcodes and NEXTflex™ DNA Barcode Blockers for SeqCap are included in the NEXTflex PreCapture Combo Kit and are also available separately.

Our whitepaper written in conjunction with Roche NimbleGen, An Integrated Solution to Simplify Library Preparation and Multiplexing for NimbleGen Sequence Capture,  demonstrates the efficacy of this kit when used with the NimbleGen SeqCap EZ Library system for library prep in preparation for Illumina sequencing.


Selected Publications that Reference Using the NEXTflex Pre-Capture Combo Library Prep Kit

Kocak, et al (2014) Hoyeraal-Hreidarsson syndrome caused by a germline mutation in the TEL patch of the telomere protein TPP1. Genes and Development. 28:2090-2102.

Koçak, H. (2014) Functional Analysis of ACD Mutations in Dyskeratosis Congenita (Unpublished doctoral dissertation). The University of Michigan.

McCarthy, SE, et. al. (2014) De novo mutations in schizophrenia implicate chromatin remodeling and support a genetic overlap with autism and intellectual disability. Mol. Psychiatry 19:652-8.

Yeh, I. et al. (2015) Activating MET kinase rearrangements in melanoma and Spitz tumours. Nature Communications. 6:7174, doi:10.1038/ncomms8174.


Kit Specs

The NEXTflex PreCapture Combo Kit (NimbleGen SeqCap EZ Library Compatible) contains DNA-seq library preparation reagents to prepare 48, 96, 192, 384 or 768 genomic DNA-seq or exome libraries for Roche NimbleGen’s SeqCap EZ Library capture systems and downstream Illumina®-compatible next-generation sequencing. The shelf life of all reagents is 12 months when stored properly at -20°C. This kit is shipped on dry ice.

NEXTflex Pre-Capture Combo Kit for Illumina Library Prep Workflow

 

NEXTflex Pre-Capture Combo Kit workflow for NimbleGen SeqCap EZ library construction and Illumina sequencing

 

 

 

Kit Contents

NEXTflex™ End Repair Buffer Mix 
NEXTflex™ End Repair Enzyme Mix 
NEXTflex™ Adenylation Mix 
NEXTflex™ Ligation Mix
NEXTflex™ DNA Barcode Adapter 1 - 6 (25 µM)
NEXTflex™ PCR Master Mix
NEXTflex™ Primer Mix (12.5 µM)
NEXTflex™ INV-HE Index 1 – 6 (250 µM)
NEXTflex™ HE Universal Oligo 1 (500 µM)
NEXTflex™ LM-PCR Oligo 1 (100 µM)
NEXTflex™ LM-PCR Oligo 2 (100 µM)
NEXTflex™ LM-PCR Master Mix (2X)
Nuclease-free Water
Resuspension Buffer

 

Required Materials not Provided

1 µg of fragmented genomic DNA in up to 40 µL nuclease-free water
NimbleGen SeqCap v3 EZ Library
All consumables required to carry out NimbleGen SeqCap v3 EZ Library
NimbleGen SeqCap EZ Hybridization and Wash Kit (Roche, Cat. # 05634253001, 96 reactions)
All consumables required to carry out NimbleGen SeqCap EZ Hybridization and Wash
Ethanol 100% (room temperature)
Ethanol 80% (room temperature)
AIR™ DNA Fragmentation Kit (Bioo Scientific, Cat # 5135-01) / or / Covaris System (S2, E210)
96 well PCR Plate Non-skirted (Phenix Research, Cat # MPS-499) / or / similar 
96 well Library Storage and Pooling Plate (Fisher Scientific, Cat # AB-0765) / or / similar
Adhesive PCR Plate Seal (BioRad, Cat # MSB1001)
Agencourt AMPure XP 60 mL (Beckman Coulter Genomics, Cat # A63881)
Magnetic Stand -96 (Ambion, Cat # AM10027) / or / similar
Heat block
Thermocycler 
2, 10, 20, 200 and 1000 L pipettes / multichannel pipettes
Nuclease-free barrier pipette tips
Microcentrifuge 
1.5 mL nuclease-free microcentrifuge tubes
Low melt agarose such as Low Gelling Temperature Agarose with a melt point of 65ºC (Boston Bioproducts, Cat # P-730)
1X TAE buffer
Clean razor or scalpel
SYBR Gold (Invitrogen, Cat # S11494)
UV transilluminator or gel documentation instrument
Gel electrophoresis apparatus
Electrophoresis power supply
Vortex

 

Sequences of NEXTflex DNA Barcode Indexes - Excel / PDF

Sequences of NEXTflex-96 DNA Barcode Indexes - Excel / PDF

Instructions for installing NEXTflex Barcode Indices in Illumina Experiment Manager


Download the whitepaper, An Integrated Solution to Simplify Library Preparation and Multiplexing for NimbleGen Sequence Capture