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RNA Library Preparation

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FAQs for RNA Library Preparation

Protocols for RNA Library Preparation

NEBNext® Magnesium RNA Fragmentation Module Protocol
Protocol for Oligonucleotide Adenylation (E2610)
Protocol for blunting ends by 3' overhang removal and fill-in of 3' recessed (5' overhang) ends using DNA Polymerase I, Large (Klenow) Fragment (M0210)
Protocol for blunting ends by 3’ overhang removal and 3’ recessed (5’ overhang) end fill-in using T4 DNA Polymerase (M0203)
Protocol for a PCR reaction using NEBNext® Q5® Hot Start HiFi PCR Master Mix (M0543)
Data Analysis - NEBNext Library Quant Kit (E7630)
Experimental Considerations - NEBNext Library Quant Kit (E7630)
NEBNext Library Quant Kit Protocol - NEBNext Library Quant Kit (E7630)
NEBNext Library Quant Kit Quick Protocol (E7630)
Expected Results - NEBNext Library Quant Kit (E7630)
Protocol for use with NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490)
Protocol for use with FFPE RNA, NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310)
Protocol for use with NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310) and NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB #E7760, #E7765)
Protocol for use with Purified mRNA or rRNA Depleted RNA and NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB #E7760, #E7765)
Protocol for use with rRNA Depleted FFPE RNA and NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB #E7760, #E7765)
Protocol for use with NEBNext® UltraTM II RNA Library Prep Kit for Illumina® E7770 and rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310)
Protocol for use with NEBNext Poly(A) mRNA Magnetic Isolation Module (E7490) and NEBNext Ultra II RNA Library Prep Kit for Illumina (E7770, E7775)
Protocol for use with FFPE RNA, NEBNext Ulta II RNA Library Prep Kit for Illumina E7770 and rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310)
Protocols for use with Purified mRNA or rRNA Depleted RNA and NEBNext Ultra II RNA Library Prep Kit for Illumina
Protocol for use with rRNA Depleted FFPE RNA and NEBNext Ultra II RNA Library Prep Kit for Illumina (NEB # E7770, #E7775)
Protocol for use with NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (E6310, E6350)
Protocol for Directional RNA-seq Workflows and NEBNext Ultra II RNA First Strand Synthesis Module (E7771)
Protocol for Non-directional RNA-seq Workflows and NEBNext Ultra II RNA First Strand Synthesis Module (E7771)
Appendix A
Appendix A for use with NEBNext Ultra II RNA First Strand Synthesis Module (E7771)

Application Notes for RNA Library Preparation

Automated size selection of NEBNext^® Small RNA libraries with the Sage Pippin Prep™

Tools & Resources

Usage Guidelines

Getting Started with RNA-Sequencing (RNA-Seq)

Legal Information

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 email us at 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.

Videos

NEBNEXT^® Ultra II Directional RNA Workflow

Learn more about the streamlined workflow for the NEBNext Ultra II Directional RNA Library Prep Kit.
Tips for Optimizing RNA Inputs in NGS Library Construction

Watch as Deyra explains how to optimize your RNA inputs for successful NGS library preparation.
Quick Tips - How can I optimize fragmentation for RNA workflows?

This video will help you optimize fragmentation for RNA workflows in the NEBNext product line.
A Breakthrough Method of Small RNA Sample Preparation for Next Gen Sequencing

The common problem of adaptor dimer formation during small RNA library construction can be avoided by using NEBNext® protocols. Learn about this technique, and how it improves both performance and sensitivity in library construction.
Biomek Automated NEBNEXT^® Ultra Directional RNA Library Prep Kit for Illumina

Learn how Biomek automation of NEBNext RNA library preparation enables high-throughput, reproducible production of libraries with high yield and complexity. In this webinar Zach Smith from Beckman Coulter describes strand-specific RNA library preparation using the NEBNext Ultra Directional RNA Library Prep Kit for Illumina and discusses Biomek automation of this method.