Product Class: Kit

NEBNext® Fast DNA Fragmentation & Library Prep Set for Ion Torrent™

The S size of this product has been discontinued as of September 2018. The L size will still be available. 

Product Introduction

The NEBNext® Fast DNA Fragmentation & Library Prep Set for Ion Torrent® includes enzymatic DNA fragmentation in a streamlined, high-performance library prep workflow.

  • High yield 
  • Fast, simple workflow – under 2 hours, with minimal hands-on time 
  • Compatible with broad range of sample input (10 ng –1 μg)
Catalog # Size Concentration
E6285L 50.0 reactions

Product Information

Description


The NEBNext® Fast DNA Fragmentation & Library Prep Set for Ion Torrent® includes a master mix of reagents for enzymatic DNA fragmentation combined with end repair. This enables efficient and tunable DNA shearing and blunting with minimized sample loss before performing adaptor ligation and high-fidelity library amplification. The kit is compatible with input amounts from 10 ng - 1µg, with a workflow time of under 2 hours and ~12 min. hands-on time.

Adaptors and primers for singleplex libraries are supplied in the kits. For multiplexed libraries, the Ion Xpress™ Barcode Adapters from Thermo Fisher Scientific can be used.

If mechanical shearing of DNA is preferred, a version of the kit without enzymatic fragmentation reagents is available.

DNA Library Preparation Workflow for Ion Torrent

DNA Library Preparation Workflow for Ion Torrent


Representative Results from Libraries Constructed Using NEBNext for Ion Torrent

Representative Results from Libraries Constructed Using NEBNext for Ion Torrent

0.5 µg of DNA from 3 different genomes with varying GC content were used to construct 200 bp and 400 bp libraries using the NEBNext Fast DNA Fragmentation and Library Prep Set for Ion Torrent, analyzed by the Agilent® Bioanalyzer®.

Representative Ion Torrent Run Report

Representative Ion Torrent Run Report
A typical Ion Torrent run report for libraries made from E. coli (K12 MG1655 strain) genomic DNA using NEBNext Fast DNA Fragmentation & Library Prep Set for Ion Torrent. A. Alignment Summary. 72 Mb of data was generated, with an average genome coverage of 15.5 X, from approximately 0.5 Million 200 bp reads. B. Read length histogram from 200 bp run.

Figure 1: Relative size distribution of Fragmented End Repaired DNA as seen using the Bioanalyzer® 2100 (Agilent Technologies, Inc.).

Figure 1: Relative size distribution of Fragmented End Repaired DNA as seen using the Bioanalyzer® 2100 (Agilent Technologies, Inc.).
1 µg of E. coli DNA was fragmented and end repaired for 20 minutes at 25°C, followed by 10 minutes at 70°C.


Figure 2: Final Library size distribution using AMPure XP Bead Size Selection.

Figure 2: Final Library size distribution using AMPure XP Bead Size Selection.



Figure 3: Final Library Size distribution using E-Gel Size Selection.

Figure 3: Final Library Size distribution using E-Gel Size Selection.

Lot Control

The lots provided are managed separately and qualified by additional functional validation. Individual reagents undergo standard enzyme activity and quality control assays, and also meet stringent criteria in the additional quality controls listed on each individual component page
This product is related to the following categories:
Library Preparation for Ion Torrent Products
This product can be used in the following applications:
DNA Sequencing

Kit Components

Kit Components

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • E6285L     -20    
      NEBNext DNA Library Primers for Ion Torrent™ E6291AAVIAL -20 1 x 0.5 ml Not Applicable
      T4 DNA Ligase E6288AAVIAL -20 1 x 0.2 ml Not Applicable
      Magnesium Chloride (MgCl2) (200 mM) E6295AAVIAL -20 1 x 0.25 ml Not Applicable
      NEBNext Stop Buffer E6294AAVIAL -20 1 x 0.25 ml Not Applicable
      NEBNext DNA Fragmentation Reaction Buffer E6287AAVIAL -20 1 x 0.1 ml Not Applicable
      NEBNext DNA Library Adaptors for Ion Torrent E6290AAVIAL -20 1 x 0.5 ml Not Applicable
      NEBNext DNA Fragmentation Master Mix E6286AAVIAL -20 1 x 0.1 ml Not Applicable
      T4 DNA Ligase Buffer for Ion Torrent E6296AAVIAL -20 1 x 0.2 ml Not Applicable
      TE Buffer E6299AAVIAL -20 1 x 7.75 ml 0.1 X
      Bst 2.0 WarmStart® DNA Polymerase E6277AAVIAL -20 1 x 0.05 ml Not Applicable
      NEBNext Q5 Hot Start HiFi PCR Master Mix E6629AAVIAL -20 2 x 1.25 ml 2 X

Properties & Usage

Materials Required but not Supplied

  • Agencourt® AMPure® XP Beads (Beckman Coulter, Inc., #A63881) or PCR Column Purification Kit (Qiagen or other)
  • 80% ethanol (freshly prepared)
  • Size selection materials [E-Gel® (Life Technologies, Inc.) or AMPURE XP Beads etc.]
  • Magnetic rack or plate (e.g., NEBNext® Magnetic Separation Rack (NEB #S1515S), Alpaqua ® 96S Super Magnet Plate (#A001322), or equivalent)

Protocols, Manuals & Usage

Protocols

  1. NEBNext® Fast DNA Fragmentation & Library Prep Set for Ion Torrent™

Manuals

The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.

FAQs & Troubleshooting

FAQs

  1. What sequencing platform can I use the NEBNext Fast DNA  Library Prep Sets for Ion Torrent reagents for?
  2. What type of starting material can be used with NEBNext Fast DNA Fragmentation & Library Prep Set for Ion Torrent?
  3. How much starting material do I need to use when preparing libraries using the NEBNext Fast DNA Fragmentation & Library Prep Set for Ion Torrent?
  4. Does the NEBNext Fast DNA Fragmentation & Library Prep Set for Ion Torrent work well for GC-rich DNA and AT-rich DNA?
  5. Can I use the NEBNext Fast DNA Fragmentation & Library Prep Set for Ion Torrent for preparing RNA?
  6. What methods for size selection can be used with Ion Torrent Library Preparation Kits?
  7. Are these reagents available in a customized format, larger format or 96 well formats?
  8. What do I do if my fragmented DNA is too short?
  9. The peak on my electropherogram trace is broader than I would like. How can I make the size range more narrow?
  10. Do I really need to vortex the fragmentation reactions, as described in the protocol? Will pipetting up and down work just as well?
  11. What methods for size selection can be used with Ion Torrent Library Preparation Kits?
  12. What do I do if my DNA is not being fragmented?
  13. Are DNA adaptors or oligonucleotides included in the NEBNext® products?
  14. Are NEBNext® products validated in Next Generation Sequencing workflows?

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

Licenses

This product is covered by one or more patents.

This product is licensed for research and commercial use from Bio-Rad Laboratories, Inc., under U.S. Pat. Nos. 6,627,424, 7,541,170, 7,670,808, 7,666,645, and corresponding patents in other countries. No rights are granted for use of the product for Digital PCR or real-time PCR applications, with the exception of quantification in Next Generation Sequencing workflows.

Nucleic acid-based aptamers for use with thermophilic DNA polymerases are licensed exclusively by New England Biolabs, Inc. from SomaLogic, Inc. New England Biolabs, Inc. gives the Buyer/User a non-exclusive license to use the NEBNext® Fast DNA Fragmentation & Library Prep Set for Ion Torrent™ for Research Use Only (RUO). Commercial use of this product may require a license from New England Biolabs, Inc. For additional information or to inquire about commercial use, please contact busdev@neb.com.

Trademarks

NEBNEXT®, NEW ENGLAND BIOLABS®, WARMSTART® and Q5® are registered trademarks of New England Biolabs, Inc.
LITMUS™ and THERMOPOL™ are trademarks of New England Biolabs, Inc.

AMPURE® is a registered trademark of Beckman Coulter, Inc.
BIOANALYZER® is a registered trademark of Agilent Technolgies, Inc.
ION XPRESS™ and ION TORRENT™ are trademarks owned by Life Technologies, Inc.

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