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

Catalog #	Size	List Price	Your Price
E7420L	96 reactions

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.

NEBNext 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 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 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 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 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)

Companion Products

NEBNext^® rRNA Depletion Kit (Human/Mouse/Rat)

Protocols

Usage Guidelines

Getting Started with RNA-Sequencing (RNA-Seq)

DataCards

manualE7420

FAQs

Troubleshooting

NEBNext® RNA Library Prep Kit Troubleshooting Guide (NEB #E7760, E7765, E7770, E7775, E7420, E7530)

Citations

See more details on Bioz

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]

E7420S_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]

Legal And Disclaimer

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.

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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