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Do you need increased sensitivity and specificity from your RNA-seq experiments? Do you have ever-decreasing amounts of input RNA? To address these challenges, our next generation of RNA library prep kits have been reformulated at each step, resulting in several fold higher yields of high quality libraries, enabling use of lower input amounts and fewer PCR cycles.

Our new NEBNext Ultra II RNA kits have streamlined, automatable workflows and are available for directional (strand-specific, using the "dUTP method") and non-directional library prep and are compatible with poly(A) mRNA enrichment or rRNA depletion. The kits are available with the option of SPRISelect® beads for size-selection and clean-up steps.

For single cell and ultra-low input RNA, our separate template-switching-based workflow generates full length cDNAs, followed by conversion to sequence-ready libraries using the Ultra II FS workflow, that incorporates enzymatic DNA fragmentation. For more information, see the NEBNext Single Cell/Low Input Library Prep Kit for Illumina.

Abundant RNA species such as rRNAs and certain mRNAs can interfere with detection and analysis of biologically relevant RNAs of interest. Several RNA depletion products are available to selectively degrade and remove unwanted RNAs from your sample. For ribosomal RNA (rRNA) depletion from human, mouse, or rat samples, use the NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310). For depletion of the abundant Globin mRNAs and rRNAs from blood samples, use the NEBNext Globin & rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E7750). For depletion of bacterial rRNA from bacterial samples, use the NEBNext rRNA Depletion Kit (Bacteria) (NEB #E7850). For oligo d(T)-based enrichment of mRNAs by binding to intact poly(A) tails, the NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) can be used.

Why consider the NEBNext Ultra II Directional RNA Library Prep Kit?

  • Get more of what you need, with the highest library yields
  • Generate high-quality libraries even with limited amounts of RNA:
    • 10 ng – 1 µg Total RNA  (poly(A) mRNA workflow)
    • 5 ng – 1 µg Total RNA (rRNA depletion workflow)
  • Minimize bias, with fewer PCR cycles required
  • Increase the complexity and transcript coverage of your libraries
  • Maximize flexibility by ordering reagents for your specific workflow needs, including directional and non-directional kits, rRNA depletion and poly(A) mRNA isolation reagents, and adaptors and primers (12- and 96-index) sold separately
  • Enjoy the reliability of the gold standard SPRIselect size selection and clean-up beads, supplied in just the amounts you need
  • Save time with streamlined workflows, reduced hands-on time, and automation compatibility
  • Rely on robust performance, even with low quality RNA, including FFPE

Are you looking for non-directional library prep?

View performance data for our NEBNext Ultra II RNA Library Prep Kit: 

The Wellcome Trust Sanger Institute has recently re-evaluated its RNA-seq library preparation methods and has chosen to implement the NEBNext Ultra II Directional RNA Library Prep Kit. Our previous RNA-seq library preparation required a minimum input of 500 ng total RNA, but the high efficiency of the NEBNext Ultra II kit has allowed us to reduce this to either 100 ng for a standard pipeline, or down to just 5 ng for a low input option, enabling RNA-seq to be carried out from just 5,000-10,000 cells.

– Mike Quail,
   Senior Staff Scientist, R & D Sequencing, Wellcome Trust Sanger Institute
 
See the full press release here.

One or more of these products are covered by patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc. For more information, please email us at gbd@neb.com. The use of these products may require you to obtain additional third party intellectual property rights for certain applications.

ILLUMINA®, TRUSEQ® and NEXTSEQ® are registered trademarks of Illumina, Inc.
SPRISELECT® is a registered trademark of Beckman Coulter, Inc.
RIBO-ZERO™ is a trademark of Illumina, Inc.
KAPA™ is a trademark of Kapa Biosystems

Available kits:

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NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina®
Includes optimized mixes for directional RNA library preparation (fragmentation, cDNA synthesis, end repair/dA-tailing, adaptor ligation and PCR enrichment steps) for sequencing on the Illumina platform.

NEBNext_portal_product_icon 

NEBNext® Ultra™ II Directional RNA Library Prep with Sample Purification Beads
Includes optimized mixes for directional RNA library preparation (fragmentation, cDNA synthesis, end repair/dA-tailing, adaptor ligation and PCR enrichment steps) plus SPRIselect® beads for size selection and cleanup.

NEBNext_portal_product_icon 

NEBNext® Ultra™ II RNA Library Prep Kit for Illumina®
Includes optimized mixes for non-directional RNA library preparation (fragmentation, cDNA synthesis, end repair/dA-tailing, adaptor ligation and PCR enrichment steps) for sequencing on the Illumina platform.

NEBNext_portal_product_icon 

NEBNext® Ultra™ II RNA Library Prep with Sample Purification Beads
Includes optimized mixes for non-directional RNA library preparation (fragmentation, cDNA synthesis, end repair/dA-tailing, adaptor ligation and PCR enrichment steps) plus SPRIselect® beads for size selection and cleanup.

Videos

NEBNext_portal_Selector_icon  NEBNext Selector
Find the right kits for your DNA or RNA sample preparation with our interactive online tool