NEBNext® Ultra™ II RNA Library Prep (Directional & Non-directional)

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.

The new NEBNext Ultra II RNA kits from New England Biolabs 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.

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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

 

NEBNext Ultra II Directional RNA produces the highest yields, from a range of input amounts

Poly(A)-containing mRNA was isolated from 10 ng, 100 ng and 1 µg of Universal Human Reference RNA (Agilent #740000) and libraries were made using the NEBNext Ultra II Directional RNA kit, Kapa Stranded mRNA-Seq kit, Kapa mRNA HyperPrep kit and Illumina TruSeq Stranded mRNA Kit. The input RNA amount and number of PCR cycles are indicated. Library yields from an average of three replicates are shown. Error bars indicate standard deviation. Library yields were assessed using the Agilent® Bioanalyzer®.

Are you looking for non-directional library prep?

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

Improving library yields

Generating high-quality libraries

Increasing library complexity

Maximizing transcript coverage

What others say about the NEBNext Ultra II RNA kit from New England Biolabs

"The Ultra II RNA kit has allowed us to reduce the input for directional polyA+ RNAseq libraries by a factor of 10 or more. We can now make a library with only 10 ng high quality total RNA and get the same gene expression profile as for 1 µg input. We've even pushed the input as low as 1 ng for very high quality total RNA. The new Ultra II RNA kit makes RNAseq achievable for low yield samples, we actually need more RNA for quality control than for library prep! Furthermore, the library prep protocol is streamlined compared to the previous Ultra RNA kits, including a reduction in AMpure bead cleanups and PCR cycles, resulting in better libraries for less time and resources."

– Jen Grenier, Ph.D.
Director of RNA Sequencing Core (RSC), Center for
Reproductive Genomics, Department of Biomedical
Sciences, College of Veterinary Medicine, Cornell University

"I used the NEBNext Ultra II Directional RNA Library Prep Kit to process very low input (7–8 ng total RNA) samples from difficult to obtain tissue for one of our customers, and I am very pleased with the results."

– Brian James, Ph.D.
Genomics Facility Director,
Sanford Burnham Prebys Medical Discovery Institute

"At The Earlham Institute we process many sample types; these include plant, microbial and animal. We trialed NEB’s Ultra II Directional RNA Library Prep using three plant species that have previously been problematic with other RNA-seq kits. We were thrilled with how well and how consistently this kit performed. The NEBNext® Poly(A) mRNA Magnetic Isolation Module was also effective and SortMeRNA showed that we had less than 2% contamination for each plant species. The strand specificity was also good at 98%. We were so pleased with the results that we are automating this protocol ready for production with in the EI Genomics Lab."

– Leah Clissold
Platforms & Pipelines Team Leader,
The Earlham Institute, Norwich, UK

"The Ultra II Directional RNA kit is very impressive... I’ve pushed the lower limit of input down to 1 and 2 ng and still achieved libraries with a good depth in complexity. Importantly, I've also noticed a reduction in adaptor-dimers and duplication in sequencing. The Ultra II is my “go-to” kit for all RNA-seq library preparations."

– Dr. S. Balu
King’s College London

 

Available kits

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® 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® 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® 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.

 

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