Product Class: Kit

NEBNext® FS DNA Library Prep Kit for MGI®

Product Introduction

The NEBNext FS DNA Library Prep Kit  for MGI is designed for use in MGI sequencing applications. This kit includes optimized components and protocols for the production of high-quality DNA libraries from a wide range (0.1 – 500 ng) of intact DNA inputs, with integrated enzymatic fragmentation, end-repair, and dA-tailing steps.

The NEBNext FS DNA Library Prep Kit is used in conjunction with NEBNext Multiplex Oligos for MGI (Dual Index Primer Pairs Set 1) (NEB #E9725) and libraries are circularized using the NEBNext Circularization Module for MGI (NEB #E9720) before sequencing.

  • Fast workflow
  • High library yields
  • Broad input range: 0.1 – 500 ng
  • Uniform GC coverage
  • Lower duplicates and chimeras
  • Minimized adaptor dimers

Request a sample of this product by visiting the NEBNext Sample Request Page and choosing “Sample not listed” at the bottom of the list; your local NEB Account Manager will reach out to confirm the details of your request.

Catalog # Size Concentration
E9705S 24.0 reactions
E9705L 96.0 reactions

Product Information

Description

The NEBNext FS DNA Library Prep Kit for MGI meets the need for reliably high-quality DNA library prep from intact samples for MGI sequencing applications.

The kit incorporates trusted NEBNext master mixed reagents and protocols, allowing the use of a wide range of intact DNA inputs (0.1 - 500 ng). The integrated enzymatic fragmentation, end-repair, and dA-tailing step offers fragment-size tunability with a single enzyme mix; the "FS" stands for Fragmentation System.   

Note that adaptors, primers, and circularization reagents are provided separately in the NEBNext Multiplex Oligos for MGI (Dual Index Primer Pairs Set 1) (NEB #E9725) and NEBNext Circularization Module for MGI (NEB #E9720).

  • Get more of what you need with high library yields
  • Generate high-quality libraries with flexible input amounts from 0.1 ng to 500 ng of intact DNA
  • Appreciate the convenience of a single adaptor-dilution recommendation
  • Optimize your time with streamlined workflows, minimized hands-on time, and automation compatibility
  • Rely on robust performance

 

FIGURE 1: NEBNext FS DNA Library Prep Kit for MGI workflow

Workflow for E9705

The NEBNext FS DNA Library Prep Kit for MGI has a streamlined workflow with minimal hands-on time across a range of inputs from 0.1 ng to 500 ng.

FIGURE 2: The NEBNext FS DNA Library Prep Kit for MGI enables robust library preparation over a wide input range without adaptor dimers

E9705 Yield

Libraries were prepared from 0.1–500 ng of human gDNA (NA12878, Coriell Institute for Medical Research) with the indicated PCR cycles. Size selection was performed for a 250 bp average fragment size, except for the 0.1 ng input, where bead cleanup was conducted to avoid excess sample loss and to preserve library complexity. Stock adaptor concentration was used for all input amounts except for 0.1 ng, where a 10-fold dilution of the stock was applied. No traces of adaptor dimer were observed (at ~150 bp) at all input levels (A). Robust library yields were obtained across input amounts. Each bar represents the average of 8–18 replicates, with error bars indicating standard deviation (B). M = Marker

FIGURE 3: The NEBNext FS DNA Library Prep Kit for MGI produces high-quality libraries with excellent sequencing metrics across the input range

E7905 Prep

Libraries were prepared in duplicate from 0.1–500 ng of NA12878 gDNA (Coriell Institute for Medical Research), sequenced on an MGI® G99 platform with PE100 and downsampled to 3.5 million paired-end reads. Reads were mapped using Bowtie2 (version 2.3.2.2) to the GRCh38 reference, and duplicates were marked using Picard MarkDuplicates (version 1.56.0). Library quality metrics were assessed using Picard Alignment Summary Metrics (version 1.56.0). High mapping rates, low chimera rates, and low adaptor-dimer rates were obtained for all libraries. A slightly higher adaptor-dimer presence (~1%) was observed with the 1 ng input for this specific experiment, and this can be optimized by adaptor dilution. Stock adaptor concentration was used for all input levels except for 0.1 ng, where a 10-fold dilution of the stock was applied.

FIGURE 4: The NEBNext FS DNA Library Prep Kit for MGI produces high-quality libraries with uniform GC coverage across the input range  

E7905 Coverage

Libraries were prepared in duplicate from 0.1–500 ng of NA12878 gDNA (Coriell Institute for Medical Research), sequenced on an MGI® G99 platform with PE100 and downsampled to 3.5 million paired-end reads. Reads were mapped using Bowtie2 (version 2.3.2.2) to the GRCh38 reference, and duplicates were marked using Picard MarkDuplicates (version 1.56.0). GC coverage was calculated using Picard's CollectGCBiasMetrics (1.56), which was run on human autosomes only, due to the even copy-number assumption of the tool. Uniform GC coverage was achieved for all input amounts. In the GC coverage plot, the horizontal grey lines indicate the expected normalized coverage of 1.0, and the dots in shades of green represent read numbers at each GC%. The grey area plots below are histograms representing the distribution of GC content in 100 bp windows of the reference genome.

FIGURE 5: The NEBNext FS DNA Library Prep Kit for MGI produces high-quality libraries with consistent insert sizes across a broad input range

E7905 Inputs

Libraries were prepared in duplicate from 0.1–500 ng of NA12878 gDNA (Coriell Institute for Medical Research) with a 250 bp size selection performed for 1–500 ng, or a simple bead cleanup step for 0.1 ng to preserve library complexity. Libraries were pooled and sequenced on an MGI G99 platform with 2x100 base reads and downsampled to 3.5 million paired-end reads. Reads were mapped using Bowtie2 (version 2.3.2.2) to the GRCh38 reference, and insert size distributions were calculated using Picard CollectInsertSizeMetrics (v1.56.0). Consistent insert sizes of 250 bp were achieved for all input amounts with size selection. In contrast, a smaller size distribution was observed for inputs of 0.1 ng where a bead-based clean-up step was used to remove adaptor dimers and excess adaptors, without size selection.

This product is related to the following categories:
Library Preparation for MGI® Products,
Next Generation Sequencing Library Preparation Products,

Kit Components

Kit Components

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • E9705S     -20    
      NEBNext® FS Reaction Buffer E9616AVIAL -20 1 x 0.168 ml Not Applicable
      NEBNext® FS Enzyme Mix E9617AVIAL -20 1 x 0.048 ml Not Applicable
      NEBNext® Ligation Master Mix E9613AVIAL -20 1 x 0.72 ml Not Applicable
      NEBNext® Ligation Enhancer E7374AVIAL -20 1 x 0.024 ml Not Applicable
      NEBNext® MSTC™ High Yield Master Mix E9615AVIAL -20 1 x 0.6 ml Not Applicable
      TE Buffer (1X) E9614AVIAL -20 1 x 1.5 ml Not Applicable
  • E9705L     -20    
      NEBNext® FS Reaction Buffer E9616AAVIAL -20 1 x 0.672 ml Not Applicable
      NEBNext® FS Enzyme Mix E9617AAVIAL -20 1 x 0.192 ml Not Applicable
      NEBNext® Ligation Master Mix E9613AAVIAL -20 3 x 0.96 ml Not Applicable
      NEBNext® Ligation Enhancer E7374AAVIAL -20 1 x 0.096 ml Not Applicable
      NEBNext® MSTC™ High Yield Master Mix E9615AAVIAL -20 2 x 1.2 ml Not Applicable
      TE Buffer (1X) E9614AAVIAL -20 1 x 6 ml Not Applicable

Properties & Usage

Materials Required but not Supplied

  • NEBNext Multiplex Oligos for MGI (Dual Index Primer Pairs Set 1) (#E9725)
  • NEBNext Circularization Module for MGI (#E9720)
  • SPRIselect™ Reagent Kit (Beckman Coulter®, Inc. #B23317) or AMPure® XP Beads (Beckman Coulter, Inc. #A63881)
  • 80% Ethanol (freshly prepared)
  • Nuclease-free Water
  • DNA LoBind® Tubes (Eppendorf® #022431021)
  • DNase-, RNase-free PCR strip tubes, for example TempAssure® PCR flex-free 8-tube strips (USA Scientific® #1402-4708)
  • Magnetic rack/stand (NEB #S1515, Alpaqua® cat. #A001322, or equivalent)
  • Vortex
  • Thermal cycler
  • Bioanalyzer® or TapeStation® (Agilent® Technologies, Inc.) and associated reagents and consumables

Protocols, Manuals & Usage

Protocols

  1. Where can I find protocols for the  NEBNext® FS DNA Library Prep kit for MGI® (NEB #E9705)?

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 do I do if I see a precipitate in the NEBNext® FS Reaction Buffer during MGI library prep?
  2. The ligation reaction is very viscous. What will happen if it is not mixed properly?
  3. Do the NEBNext® Library Prep kits for MGI® include beads?
  4. Which kit can I use for circularization to make single-stranded circular DNA libraries for sequencing on MGI® platforms?
  5. Can the NEBNext® FS DNA Library Prep Kit for MGI® (NEB #E9705) provide sufficient library yield for target enrichment?
  6. How much library is required for the circularization reaction when using the NEBNext® Circularization Module for MGI® (NEB #E9635)?
  7. When preparing samples using the NEBNext® FS DNA Library Prep Kit for MGI® (NEB #E9705), can my input DNA be in EDTA-containing solutions?
  8. How much starting material must I use when preparing libraries with the NEBNext® FS DNA Library Prep Kit for MGI® (NEB #E9705)?
  9. Can I use the NEBNext® FS DNA Library Prep Kit for MGI® (NEB #E9705) for bisulfite conversion and EM-seq™ workflows?
  10. Do I need to vortex the NEBNext® FS Enzyme Mix?
  11. What sample types can I use with the NEBNext® FS DNA Library Prep Kit for MGI® (NEB #E9705)?
  12. Do the NEBNext® library prep kits for MGI® include adaptors and primers?
  13. Can the NEBNext® RNA Library Prep Kit for MGI® (NEB #E9710) and NEBNext FS DNA Library Prep Kit for MGI (NEB #E9705) be used with adaptors and primers from suppliers other than NEB®?

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.

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