RNA for Illumina®

For use with the Illumina® sequencing platform, RNA libraries are prepared by depletion of abundant RNAs from total RNA, and synthesis of cDNA followed by DNA library preparation steps: repair of 3′ and 5′ ends followed by the addition of a non-templated dA-tail before ligation with an adaptor. Libraries are size selected after adaptor ligation, and amplified via polymerase chain reaction (PCR). For more information, choose either the Ultra™ II Directional or Non-directional RNA Library Preparation Workflow for Illumina tab below.

Abundant RNAs can be removed from total RNA directly using an RNA depletion product specific for the sample species being used, or by enrichment of mRNA by a poly(A)-based method.

For single cell and ultra-low input RNA samples, libraries are constructed with a template-switching method to generate full-length cDNAs, followed by Ultra II library preparation. For more information, choose the Single Cell/Low Input RNA Library Preparation Workflow for Illumina tab below.

Small RNA libraries are constructed using a different workflow, in which adaptors are ligated directly to the small RNA molecules, followed by reverse transcription, PCR amplification and size selection. In the novel NEBNext® workflow, the introduction of the RT primer ahead of ligation of the 5’ adaptor prevents formation of contaminating adaptor-dimers. For more information, choose the Small RNA Library Preparation Workflow for Illumina tab below.

NEBNext reagents are available for each step in RNA and small RNA library preparation workflows for the Illumina platform. Reagents from NEB are available in multiple formats, and a wide range of index primers for multiplexing is available. Each of these reagents has been functionally validated by library preparation followed by Illumina sequencing.

Illumina® is a registered trademark of Illumina, Inc.

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FAQs for RNA for Illumina®
Protocols for RNA for Illumina®
Single Cell/Low Input RNA Library Preparation Workflow for Illumina
Workflow"
Ultra II Non-directional RNA Library Preparation Workflow for Illumina
Ultra II Directional RNA Library Preparation Workflow for Illumina
RNA Product Details for Illumina
Small RNA Library Preparation Workflow for Illumina
Small RNA Product Details for Illumina
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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 contact NEB's Global Business Development team at [email protected].

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.