NEBNext® Library Quant Kit for Illumina®

Catalog #	Concentration	Size
E7630S	Not Applicable	100 reactions
E7630L	Not Applicable	500 reactions
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Typical results from the NEBNext Library Quant Kit on a Bio-Rad CFX96 Touch (A) and an Applied Biosystems 7500 Fast real-time qPCR instrument (B). Amplification curves are shown on the left and resulting standard curves on the right. All default settings were used on both platforms. ABI 7500 Fast assay included 1X ROX (Low Concentration) and the ROX normalization.

The NEBNext Library Quant Kit components have been optimized to deliver significant improvements to qPCR-based library quantitation for Illumina sequencing. The kit contains primers which target the P5 and P7 Illumina adaptor sequences, and a set of four high-quality, pre-diluted DNA standards to enable reliable quantitation of diluted DNA libraries between 150–1000 bp.

Features

Provides more accurate and reproducible quant values than alternative methods and kits
Compatible with libraries with a broad range of insert sizes and GC content, made by a variety of methods
Requires only 4 standards, allowing more libraries to be quantitated per kit
Supplied with a convenient Library Dilution Buffer
The NEBNext Library Quant Master Mix requires only the addition of primers
Utilizes a single extension time for all libraries, regardless of insert size
Library quant values can be easily calculated using NEB’s online tool, at NEBioCalculator.neb.com
ROX is included in the kit, for use with qPCR instruments that require a reference dye for normalization

Each set of reagents is functionally validated together through qPCR-based library quantitation assays. Reagents pass the functional test by adhering to stringent criteria set for assay efficiency and quantitation cycles (Cq) for each DNA standard and no template control reaction.

For larger volume requirements, customized and bulk packaging is available by purchasing through the OEM/Bulks department at NEB. Please contact [email protected] for further information.

qPCR provides more consistent library quantitation results than Bioanalyzer® analysis — Concentrations of 4 libraries were determined by the NEBNext Library Quant Kit (orange) and compared to values measured using the Aglient Bioanalyzer (blue). Compared to NEBNext’s qPCR-based method, the Bioanalyzer concentrations displayed a greater level of variation.

Greater reproducibility of library quantitation with the NEBNext Library Quant Kit — Three 340–400 bp libraries were quantitated by 4 different users 2–4 times using either the NEBNext or KapaTM Library Quantification Kit (Universal). A notable improvement in quantitation consistency was observed for concentrations determined by the NEBNext Kit (orange) versus those from the Kapa kit (gray).

Greater lot-to-lot consistency of standards with the NEBNext Library Quant Kit — Accurate qPCR quantitation requires the use of high-quality DNA standards with known concentrations. The NEBNext Library Quant Kit contains 4 standards produced with a high level of both quantitation accuracy and consistency. This figure shows data from >70 total runs from 4 lots of both NEBNext (orange) and Kapa (gray) standards, with all C_q values plotted. Box and whiskers indicate mean and quartiles. The NEBNext Library Standards displayed much lower variation in C_q, resulting in more consistent quantitation performance.

The NEBNext Library Quant Kit values enable optimal cluster densities — Seven different libraries were quantitated using either the NEBNext Library Quant Kit (orange) or the Kapa Library Quantification Kit (Universal) (gray). Undiluted library concentrations ranged from 2–200 nM. Libraries were diluted to 8 pM and loaded onto a MiSeq® instrument (v2 chemistry; MCS v2.4.1.3). Libraries quantitated with the NEBNext kit resulted in a raw cluster density average of 1160 k/mm2, directly in the optimal range of 900–1300 k/mm2. In contrast, libraries loaded based on the Kapa quantitation averaged only 660 k/mm2.

Accurate Library Quantitation is achieved with a broad range of library size and GC content — A selection of libraries successfully quantitated with the NEBNext Library Quant Kit. Libraries are plotted by size, ranging from smaller libraries (sRNA, FFPE) at 150–230 bp to largest libraries at 980 bp. Various input sources were used, ranging from 20-70% GC content, as indicated by text color. Libraries were prepared using NEBNext, Illumina TruSeq® Nano and Kapa Hyper library prep kits (data not shown). No dependence on size or GC content was observed in library quantitation when using NEBNext.

Reagents Supplied

The following reagents are supplied with this product:

	Store at (°C)	Concentration
NEBNext Library Quant Master Mix	-20
NEBNext Library Quant Primer Mix	-20
NEBNext® Library Dilution Buffer	-20	10 X
NEBNext Library Quant DNA Standard 1
NEBNext Library Quant DNA Standard 2	-20
NEBNext Library Quant DNA Standard 3	-20
NEBNext Library Quant DNA Standard 4	-20
ROX (Low)	-20
ROX (High)	-20

Product Categories:: Library Quantitation Products,; ChIP-Seq Library Prep Products,; FFPE DNA Products; Microbiome DNA Enrichment Products; DNA Library Prep Products; RNA Library Prep Products

Applications:: RNA Library Preparation,; DNA Library Preparation,; Illumina Library Preparation

Advantages and Features
Features
- Provides more accurate and reproducible quant values than alternative methods and kits
- Compatible with libraries with a broad range of insert sizes and GC content, made by a variety of methods
- Requires only 4 standards, allowing more libraries to be quantitated per kit
- Supplied with a convenient, Library Dilution Buffer
- The NEBNext Library Quant Master Mix requires only the addition of primers
- Utilizes a single extension time for all libraries, regardless of insert size
- Library quant values can be easily calculated using NEB’s online tool, at NEBioCalculator.neb.com
- ROX is included in the kit, for use with qPCR instruments that require a reference dye for normalization
Properties & Usage
Materials Required but not Supplied
- Nuclease-free water
- qPCR machine
- qPCR plates and seals
- PCR strip tubes or microcentrifuge tubes
- Conical centrifuge tubes
Storage Temperature
-20°C
Related Products
Companion Products
- NEBNext® Library Dilution Buffer
Materials Sold Separately
- NEBNext® Library Dilution Buffer
Product Notes
1. Storage of Kit Components:
  All of the individual components of the NEBNext Library Quant Kit for Illumina are stable at –20°C for two years. When not in use, kit components should be stored at –20°C. All components of the kit, including the combined NEBNext Library Quant Master Mix and Primer Mix, are stable for at least 30 freeze/thaws.
  
  Once the Primer Mix has been added to the NEBNext Library Quant Master Mix (see Protocol on page 6 of manual), the resulting mix can be stored at –20°C for seven months. For short-term storage, this mix may be stored at 4°C for up to six weeks.
  
  Reactions can be prepared at room temperature, but unused portions of the kit reagents should be kept on ice after thawing them for use.
2. For Reagents Supplied:
  
  If you need dilution buffer in addition to what is provided in the kit, it can be purchased separately.

Protocols, Manuals & Usage
- Protocols
  NEBNext Library Quant Kit Quick Protocol (E7630)
  
  Experimental Considerations - NEBNext Library Quant Kit (E7630)
  
  NEBNext Library Quant Kit Protocol - NEBNext Library Quant Kit (E7630)
  
  Data Analysis - NEBNext Library Quant Kit (E7630)
  
  Expected Results - NEBNext Library Quant Kit (E7630)
- Manuals
  The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.
  
  manualE7630
- Application Notes
  Improved library quantitation for a broad range of library types E7630
FAQs & Troubleshooting
- FAQs
  What are the advantages of the qPCR-based NEBNext Library Quant Kit compared to other methods of quantitating NGS libraries?
  
  What libraries can be quantitated with the Quant Kit?
  
  How much should I dilute my library to run in the Quant Kit?
  
  Can I run the NEBNext Library Quant Kit on my qPCR machine?
  
  Which ROX should I use for my qPCR machine?
  
  Are 4 standards enough for a qPCR quantitation?
  
  Does the library quant require a size correction?
  
  What other materials/information do I need to run the kit?
  
  How many libraries can I quantitate at a time?
  
  How long is a run of the Quant Kit?
  
  Do you provide any analysis tools to help with the quantitation?
  
  Can I see adaptor dimers as a measure of library quality?
  
  How long can I store the master mix after adding the primers?
  
  Can the NEBNext Library Dilution Buffer be purchased separately?
Citations & Technical Literature
- Citations
  
  Product Citation Tool
  
  Powered by Bioz See more details on Bioz
- Feature Articles
  The Quantitation Question: How does accurate library quantitation influence sequencing?
Quality, Safety & Legal
- Quality Control Assays
  
  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]
  
  E7630S_L_v1
- 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]
  
  E7630L_v0_10010901
  
  E7630S_v0_10011217
  
  E7630S_v0_10013445
  
  E7630S_v1_10011217
  
  E7630L_v1_10016519
  
  E7630S_v1_10017620
  
  E7630L_v1_10019879
  
  E7630S_v1_10019924
  
  E7630L_v1_10023912
  
  E7630L_v1_10028725
  
  E7630S_v1_10026838
  
  E7630L_v1_10033111
  
  E7630S_v1_10032067
  
  E7630S_v1_10035302
  
  E7630L_v1_10036419
  
  E7630L_v1_10036443
  
  E7630L_v1_10039716
  
  E7630S_v1_10038647
  
  E7630L_v1_10027352
  
  E7630S_v1_10042942
  
  E7630L_v1_10045807
  
  E7630L_v1_10046598
  
  E7630S_v1_10047426
  
  E7630L_v0_10050075
  
  E7630S_v1_10050511
  
  E7630L_v1_10051827
  
  E7630L_v1_10042112
- Safety Data Sheets
  The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
  
  NEBNext Library Quant Master Mix
  
  NEBNext Library Quant Primer Mix
  
  NEBNext® Library Dilution Buffer
  
  NEBNext Library Quant DNA Standard 1
  
  NEBNext Library Quant DNA Standard 2
  
  NEBNext Library Quant DNA Standard 3
  
  NEBNext Library Quant DNA Standard 4
  
  ROX (Low)
  
  ROX (High)
- Legal and Disclaimers
  
  This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).
  
  While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
  
  For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].
  
  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.

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You’ll be Thrilled to Pieces

Tools

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NEBNext^® Library Quant Kit for Illumina^®

Reagents Supplied

Features

Materials Required but not Supplied

Storage Temperature

Companion Products

Materials Sold Separately

Product Citation Tool

NEBNext® Library Quant Kit for Illumina®

Reagents Supplied

Features

Materials Required but not Supplied

Storage Temperature

Companion Products

Materials Sold Separately

Product Citation Tool

Ineligible item added to cart

NEBNext^® Library Quant Kit for Illumina^®