NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina®

Catalog # Concentration Size List Price Quantity Your Price
E7420L 96 reactions
Please Inquire
Catalog # Size List Price Your Price
E7420L 96 reactions
Please Inquire
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
This original version of the Ultra Directional RNA Library Prep Kit enables strand-specific RNA library preparation in a streamlined workflow. However, we now recommend the new NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB #E7760), which has been further optimized to provide increased yields of high quality libraries from a broader range of input amounts.
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The NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina® contains reagents for preparation of strand-specific RNA libraries for Illumina sequencing. Please note that adaptors, primers, rRNA depletion reagents and poly(A) mRNA isolation reagents 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 Directional RNA Library Preparation for IlluminaUltra Direct

NEBNext Ultra™ Directional RNA produces libraries with high directionality/strand specificityNEBNext Ultra™ Directional RNA produces libraries with high directionality/strand specificity

Ultra Directional RNA libraries of mRNA purified from Universal Human Reference RNA (Agilent® #740000), were sequenced on the Illumina platform. The high percentage of reads mapped to the expected strand shows the high strand-specificity of the libraries. Actinomycin D is required to achieve maximal strand specificity.

NEBNext Ultra™ Directional RNA libraries provide even coverage across transcriptsNEBNext Ultra™ Directional RNA libraries provide even coverage across transcripts

mRNA was isolated from Universal Human Reference RNA (Agilent # 740000) and libraries were made using the NEBNext Ultra™ Directional RNA Kit and the Illumina TruSeq Stranded mRNA Kit. Libraries were sequenced (single end reads) on the Illumina platform (GAIIx). Good coverage across expressed transcripts was obtained.

Low input NEBNext Ultra Directional RNA libraries retain complexityLow input NEBNext Ultra Directional RNA libraries retain complexity

Ribosomal RNA-depleted RNA libraries were made from 10 ng or 100 ng Universal Human Reference RNA (Agilent #740000) using the NEBNext Ultra Directional RNA Kit. Transcript abundances in Fragments Per Kilobase of exon per Million fragments mapped (FPKM) show strong concordance (R2 = 0.90745), indicating retention of library complexity even at low input amount.

Equivalent transcript representation is achieved between technical replicates of NEBNext Ultra Directional RNA librariesEquivalent transcript representation is achieved between technical replicates of NEBNext Ultra Directional RNA libraries

Technical replicates of mRNA libraries made from mRNA isolated from Universal Human Reference RNA (Agilent #740000) show strong concordance (R2 = 0.99409).


Sequencing data shows strong concordance between NEBNext Ultra Directional RNA and Illumina TruSeq Stranded RNA librariesSequencing data shows strong concordance between NEBNext Ultra Directional RNA and Illumina TruSeq Stranded RNA libraries

mRNA was isolated from Universal Human Reference RNA (Agilent #740000) and libraries were made using the NEBNext Ultra Directional RNA Kit and the Illumina TruSeq Stranded mRNA Kit. This FPKM concordance plot shows comparable transcript representation (R2 = 0.96009) between the libraries.
Features
  • Accurate retention of transcript strand of origin information
  • Utilizes dUTP-based methodology
  • Low input amounts (as low as 10ng purified mRNA or ribosomal-depleted RNA, or 100ng Total RNA)
  • Fast workflow (4.5 – 5 hours), with minimal hands-on time
  • Robust, reliable performance
  • Ultra high fidelity amplification with minimized GC bias

Properties & Usage

Materials Required but not Supplied

  • NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) or NEBNext rRNA Depletion Kit (NEB #E6310)
  • NEBNext Singleplex (NEB #E7350) or NEBNext Multiplex (NEB #E7335, #E7500, #E7710, #E7730, #E6609#E7600) Oligos for Illumina or customer supplied Oligos
  • Magnetic rack or plate (e.g., NEBNext® Magnetic Separation Rack (NEB #S1515S), Alpaqua ® 96S Super Magnet Plate (#A001322), or equivalent)
  • 80% Ethanol (freshly prepared) 
  • 0.1X TE, pH 8.0
  • 10 mM Tris-HCl, pH 7.5–8.0
  • 10 mM NaCl (optional)
  • Agencourt® AMPure® XP Beads (Beckman Coulter, Inc. #A63881)
  • Actinomycin D (Sigma# A1410, dissolved in dimethylsulfoxide [DMSO] to 5 μg/ul)
  • Additional required materials not included for use with NEBNext rRNA Depletion Kit (NEB #E6310): Agencourt RNAClean® XP (Beckman Coulter, Inc., Cat #A63987)


Quality Control Assay
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]
Legal And Disclaimer

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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.

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.

This product and its use are the subject of one or more issued and/or pending U.S. and foreign patent applications owned by Max Planck Gesellschaft. The purchase of this product from New England Biolabs, Inc., its affiliates, or its authorized resellers and distributors conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for profit entity) or for limited commercial purposes that are pre-approved by New England Biolabs, Inc. The purchase of this product does not convey a license under any claims in the foregoing patents or patent applications directed to producing the product. The buyer cannot sell or otherwise transfer this product or its components to a third party or otherwise use this product for the following COMMERCIAL PURPOSES: (1) use of the products or its components in manufacturing; (2) use of the product or its components for therapeutic or prophylactic purposes in humans or animals.

For additional information or to inquire about commercial use, please contact busdev@neb.com.

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