Product Class: Other

NEBNext® Library Quant Kit for Ultima Genomics®


Catalog #E3410

Product Introduction

The NEBNext® Library Quant Kit for Ultima Genomics® contains the reagents necessary for qPCR-based quantitation of NGS libraries prepared using the UG 100 SolarisTM Free workflow. The kit includes standards and primers specifically developed to quantitate molecules containing the Ultima Genomics adaptor sequences, ensuring unparalleled accuracy and reliability. The streamlined protocol simplifies the reaction setup, enhancing efficiency and ensuring consistent, high-quality results for pooling Ultima Genomics Solaris Free compatible libraries.

  • Achieve sensitive and reproducible quantitation of libraries across a wide concentration range
  • Attain precision with 5 pre-diluted standards and primers optimized for Ultima Genomics Solaris Free compatible libraries
  • Simplify your reaction setup with fewer pipetting steps and a single extension time for all libraries
  • Skip the addition of ROX dye for normalization

Product Information

Description

The NEBNext Library Quant Kit for Ultima Genomics delivers reliable and streamlined qPCR-based quantitation for Ultima Genomics Solaris Free compatible libraries. The kit includes primers specific to Ultima Genomics adaptor sequences, and a set of five high-quality, pre-diluted DNA standards to enable reliable quantitation of diluted DNA libraries.

Each set of reagents is functionally validated with qPCR-based library quantitation assays. Reagents must 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.

Figure 1. NEBNext® Library Quant Kit for Ultima Genomics® workflow
Workflow for NEBNext Library Quant Kit 

 

Figure 2. Typical amplification curves
Typical amplification curves and standard curves for Bio-Rad® CFX96 Touch™ and Applied Biosystems® 7500 Fast
Typical amplification curves for the NEBNext® Library Quant Kit for Ultima Genomics®. Typical results from the NEBNext® Library Quant Kit for Ultima Genomics® on a Bio-Rad® CFX96 Touch™ (A) and an Applied Biosystems® 7500 Fast (B) real-time qPCR instrument. Amplification curves for standards 1-5 (Std 1-5) and no-template control (NTC) are shown on the left and resulting standard curves with linear fit on the right. Default settings for SYBR® Green based quantitation were used on both platforms. All standards were assayed in triplicates. Linear fit for the data provided an R2 value of ≥ 0.998 demonstrating excellent linearity of the standards with an efficiency of 90%.

 

Figure 3. Tighter distribution of sequencing reads with qPCR-based pooling
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qPCR-based pooling of libraries results in a tighter distribution of sequencing reads compared to volume-based pooling. PCR-free libraries generated from 7 different DNA input amounts (1 µg, 0.6 µg, 0.5 µg, 0.4 µg, 0.3 µg, 0.25 µg and 0.2 µg) were pooled based on either volume or qPCR quantitation and sequenced on two UG 100TM wafers. Box plots show the distribution of reads per library. Pooling based on qPCR quantitation demonstrates a much tighter distribution of reads across input amounts compared to volume-based pooling.
This product is related to the following categories:
NGS Library Quantitation,
Next Generation Sequencing Library Preparation,

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • E3410S     -20    
      Luna® Universal qPCR Master Mix E3409AVIAL -20 1 x 0.5 ml Not Applicable
      NEBNext® UG Library Quant Standard 1 E3411AVIAL -20 1 x 0.01 ml 30 pM
      NEBNext® UG Library Quant Standard 2 E3412AVIAL -20 1 x 0.01 ml 3 pM
      NEBNext® UG Library Quant Standard 3 E3413AVIAL -20 1 x 0.01 ml 0.3 pM
      NEBNext® UG Library Quant Standard 4 E3414AVIAL -20 1 x 0.01 ml 0.03 pM
      NEBNext® UG Library Quant Standard 5 E3415AVIAL -20 1 x 0.01 ml 0.003 pM
      NEBNext® UG Library Quant Primer Mix E3416AVIAL -20 1 x 0.32 ml Not Applicable
      NEBNext® Library Dilution Buffer E7633AVIAL -20 1 x 1 ml 10 X

Properties & Usage

Features

  • Developed in collaboration with Ultima Genomics
  • Compatible with libraries with a range of insert sizes, GC contents, from a variety of preparation methods
  • Uses a single extension time for all libraries, irrespective of their size
  • Visible tracking dye serves as an internal reference (no ROX required) and minimizes pipetting errors
  • Automation friendly
  • Library quant values easily calculated using NEB’s online tool, at NEBioCalculator.neb.com

Protocols, Manuals & Usage

Protocols

  1. Where can I find protocols for using the NEBNext® Library Quant Kit for Ultima Genomics® (NEB #E3410)?

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 is the NEBNext® Library Quant Kit for Ultima Genomics® (NEB #E3410)?
  2. What are the advantages of the qPCR-based NEBNext® Library Quant Kit for Ultima Genomics® (NEB #E3410) compared to other methods of quantitating NGS libraries?
  3. How much should I dilute my library to run in the NEBNext® Library Quant Kit for Ultima Genomics® (NEB #E3410)?
  4. What is the workflow time for the NEBNext® Library Quant Kit?
  5. Do you provide any analysis tools to help with the quantitation when using the NEBNext® Library Quant Kit for Ultima Genomics® (NEB #E3410)?
  6. Does the library quant require a size correction when using the NEBNext® Library Quant Kit for Ultima Genomics® (NEB #E3410)?
  7. How many libraries can I quantitate at a time when using the NEBNext® Library Quant Kit for Ultima Genomics® (NEB #E3410)?
  8. What other materials/information do I need to run an NEBNext® Library Quant Kit?
  9. Do I need to add ROX when using the NEBNext® Library Quant Kit for Ultima Genomics® (NEB #E3410)?
  10. Can I run the NEBNext® Library Quant Kit for Ultima Genomics® (NEB #E3410) on my qPCR machine?
  11. Is the NEBNext® Library Quant Kit for Ultima Genomics® (NEB #E3410) compatible with Solaris Free and Flex workflows on the Ultima Genomics platform?

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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