NEBNext® Ultra™ DNA Library Prep Kit for Illumina®
The small size of this product was discontinued on December 15, 2021. If you would like to purchase a small size, an alternative is NEB #E7645S.
This original version of the Ultra DNA Library Prep Kit enables library preparation from 5 ng – 1 µg of input DNA in a streamlined workflow. However, we now recommend use of the more recent NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB #E7645), which has been further optimized to provide increased yields of high quality libraries from a broader range of input amounts (500 pg – 1 µg).
The NEBNext® Ultra™ DNA Library Prep Kit for Illumina® contains reagents for preparation of libraries for next-generation sequencing on the Illumina platform from 5 ng – 1 µg input DNA, in a streamlined workflow. Please note that adaptors and primers are not included in the kit and are available separately.
Each kit component must pass rigorous quality control standards, and each set of reagents is functionally validated together by construction and sequencing of a library on the Illumina sequencing platform.
Ultra DNA Library Preparation Workflow for Illumina NEBNext Ultra™ DNA provides high library yields even with low inputs and difficult samples
All libraries were prepared with 5 ng of input DNA and run on an Agilent Bioanalyzer®.
A. E. coli genomic DNA libraries. 1: Ladder; 2: Library prepared using NEBNext DNA Library Prep Master Mix Set (E6040); 3: Library prepared using NEBNext Ultra DNA Library Prep Kit (E7370). With 5 ng inputs, library yields are significantly higher using the NEBNext Ultra™ DNA Kit.
B. FFPE sample. Human lung tumor genomic DNA from a 66-year old male, extracted from a 10-year old FFPE block (Biochain, Inc.). 1: Library prepared using NEBNext Ultra DNA Library Prep Kit (E7370). 2: Input DNA (highly degraded FFPE genomic DNA).Read depth correlation shows consistent high coverage for 5 ng - 1 µg input amounts Libraries were prepared with various amounts of human IMR90 gDNA (5, 50, and 1000 ng) and PCR amplified. A PCR-free library was also prepared with 1000 ng human IMR90 gDNA. Sequence coverage of 10 kb windows of the human genome was analyzed on Galaxy, and Pearson correlation coefficient (R2 values) were calculated using Microsoft® Excel®. All libraries are highly correlated with each other, independent of DNA input as well as PCR amplification. High coverage was achieved for all DNA input amounts.
NEBNext Ultra DNA Kits display strong performance with input amounts as low as 5 ng
Input DNA: IMR90 genomic DNA (human fibroblast).
PF Mismatch Rate: The rate of bases mismatching the reference for all bases aligned to the reference sequence.
Percent Duplication: The percentage of mapped sequence that is marked as duplicate.
Percent Chimeras: The percentage of reads that map outside of a maximum insert size or that have the two ends mapping to different chromosomes.
This data illustrates the strong performance of the NEBNext Ultra DNA Library Prep Kit for Illumina, even with 5 ng of human genomic DNA input. Mismatch rate, % duplication and % chimeras are all low, and similar, regardless of input amount.
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
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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 & Change Notifications
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]
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]
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.
For additional information or to inquire about commercial use, please contact busdev@neb.com.
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