产品信息

The Ultra II Directional RNA Library Prep Kit for Illumina delivers significantly increased sensitivity and specificity from your RNA-seq experiments, from ever-decreasing amounts of input RNA. In conjunction with ribosomal RNA (rRNA) depletion or poly(A) enrichment, the kit enables the production of high quality libraries from 10 ng of Total RNA, respectively, up to 1 µg.

Strand-specific/directional methods for sequencing RNA provide information on the DNA strand from which the RNA strand was transcribed. This is useful for many reasons including: Identification of antisense transcripts, determination of the transcribed strand of noncoding RNA, and measurement of expression levels of coding or noncoding overlapping transcripts. Overall, the ability to determine the originating strand can substantially enhance the value of a RNA-seq experiment.

The NEBNext Ultra II Directional RNA Library Prep Kit derives its directionality from the “dUTP” method for strand-specificity, with proven superiority for this application.

See what customers are saying about NEBNext Ultra II Directional RNA.

Features

• Get more of what you need, with the highest library yields
• Generate high quality libraries even when you have only limited amounts of input RNA:
  
  • 10 ng – 1 µg Total RNA (polyA mRNA workflow)
  • 10 ng – 1 µg Total RNA (rRNA depletion workflow)
• Minimize bias, with fewer PCR cycles required
• Increase the complexity and transcript coverage of your libraries
• Optimize your time with streamlined workflows, reduced hands-on time, and automation compatibility
• Rely on robust performance, even with low quality RNA, including FFPE
• Compatible with NEBNext poly(A) mRNA Isolation, rRNA Depletion reagents and multiplexing adaptors and primers.

Also available with optional SPRIselect® beads for clean-up and size-selection steps.

Please note that adaptors, primers, rRNA depletion reagents and poly(A) mRNA isolation reagents are not included in the kit and are available separately.

For extensive NEBNext Ultra II performance data, click the links in the Features above and download our technical note for poly(A) mRNA isolation or our technical note for rRNA depletion.

For use with NEBNext Multiplex Oligos for Illumina (Unique Dual Index UMI Adaptors RNA Set 1) (NEB #E7416), refer to the Protocols tab for UMI Adaptors-specific guidance.

LIBRARY YIELDS

Figure 1. NEBNext Ultra II Directional RNA produces the highest yields, from a range of input amounts

View additional data on library yields.

GC CONTENT DISTRIBUTION

Figure 2. NEBNext Ultra II Directional RNA libraries provide uniform GC content distribution, at a broad range of input amounts

View additional data on library quality.

MAXIMIZING TRANSCRIPT COVERAGE

Figure 3. NEBNext Ultra II Directional RNA libraries provide uniform coverage across the gene body of transcripts

View additional data on transcript coverage.

SUPERIOR LIBRARY COMPLEXITY AT LOW INPUT AMOUNTS

Figure 4. Low input NEBNext Ultra II Directional RNA libraries retain superior complexity

View additional data on library complexity.

SUPERIOR PERFORMANCE WITH FFPE RNA

Figure 5. NEBNext Ultra II Directional RNA with NEBNext rRNA Depletion results in the lowest remaining ribosomal RNA levels with FFPE samples

Figure 6. Uniformity of Coverage across the AP000769.1-201 transcript

View additional data on FFPE RNA samples.

产品类别:

Automation for NEBNext® NGS Library Prep Products,

RNA Library Prep for Illumina,

Next Generation Sequencing Library Preparation Products

应用:

Applications of USER® and Thermolabile USER II Enzymes

• 试剂盒组成

  本产品提供以下试剂或组分：

  NEB #	名称	组分货号	储存温度	数量	浓度
  E7760S     -20       NEBNext® Ligation Enhancer E7374AVIAL -20 1 x 0.024 ml Not Applicable   NEBNext First Strand Synthesis Reaction Buffer E7421AVIAL -20 1 x 0.192 ml Not Applicable   Random Primers E7422AVIAL -20 1 x 0.048 ml Not Applicable   NEBNext Second Strand Synthesis Enzyme Mix E7425AVIAL -20 1 x 0.096 ml Not Applicable   NEBNext Second Strand Synthesis Reaction Buffer with dUTP Mix E7426AVIAL -20 1 x 0.192 ml Not Applicable   NEBNext USER® Enzyme E7428AVIAL -20 1 x 0.072 ml Not Applicable   NEBNext® Ultra II End Prep Enzyme Mix E7646AVIAL -20 1 x 0.072 ml Not Applicable   NEBNext® Ultra II End Prep Reaction Buffer E7647AVIAL -20 1 x 0.168 ml Not Applicable   NEBNext® Ultra™ II Ligation Master Mix E7648AVIAL -20 1 x 0.72 ml Not Applicable   NEBNext® Ultra II Q5® Master Mix E7649AVIAL -20 1 x 0.6 ml Not Applicable   NEBNext First Strand Synthesis Enzyme Mix E7761AVIAL -20 1 x 0.048 ml Not Applicable   NEBNext® Adaptor Dilution Buffer E7762AVIAL -20 1 x 2.4 ml Not Applicable   (0.1X) TE Buffer E7763AVIAL -20 1 x 2.78 ml Not Applicable   Nuclease-free Water E7764AVIAL -20 1 x 1.25 ml Not Applicable   NEBNext Strand Specificity Reagent E7766AVIAL -20 1 x 0.192 ml Not Applicable
  E7760L     -20       NEBNext® Ligation Enhancer E7374AAVIAL -20 1 x 0.096 ml Not Applicable   NEBNext First Strand Synthesis Reaction Buffer E7421AAVIAL -20 1 x 0.768 ml Not Applicable   Random Primers E7422AAVIAL -20 1 x 0.192 ml Not Applicable   NEBNext Second Strand Synthesis Enzyme Mix E7425AAVIAL -20 1 x 0.384 ml Not Applicable   NEBNext Second Strand Synthesis Reaction Buffer with dUTP Mix E7426AAVIAL -20 1 x 0.768 ml Not Applicable   NEBNext USER® Enzyme E7428AAVIAL -20 1 x 0.288 ml Not Applicable   NEBNext® Ultra II End Prep Enzyme Mix E7646AAVIAL -20 1 x 0.288 ml Not Applicable   NEBNext® Ultra II End Prep Reaction Buffer E7647AAVIAL -20 1 x 0.672 ml Not Applicable   NEBNext® Ultra™ II Ligation Master Mix E7648AAVIAL -20 3 x 0.96 ml Not Applicable   NEBNext® Ultra II Q5® Master Mix E7649AAVIAL -20 2 x 1.2 ml Not Applicable   NEBNext First Strand Synthesis Enzyme Mix E7761AAVIAL -20 1 x 0.192 ml Not Applicable   NEBNext® Adaptor Dilution Buffer E7762AAVIAL -20 1 x 9.6 ml Not Applicable   (0.1X) TE Buffer E7763AAVIAL -20 1 x 11.5 ml Not Applicable   Nuclease-free Water E7764AAVIAL -20 1 x 5.7 ml Not Applicable   NEBNext Strand Specificity Reagent E7766AAVIAL -20 1 x 0.768 ml Not Applicable

• 特性和用法

  需要但不提供的材料

  • NEBNext Singleplex or Multiplex Oligos for Illumina (NEB.com/oligos) or customer supplied oligos
  • Magnetic rack or plate (e.g., NEBNext^® Magnetic Separation Rack (NEB #S1515S), Alpaqua ^® 96S Super Magnet Plate (#A001322), or equivalent)
  • SPRIselect® Reagent Kit (Beckman Coulter, Inc. #B23317) or AMPure® XP Beads (Beckman Coulter, Inc. #A63881)
  • 80% Ethanol (freshly prepared)
  • Thermal Cycler
  • NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) 
    or NEBNext^® rRNA Depletion Kit v2 (Human/Mouse/Rat) (NEB #E7400 or NEB #E7405)
    or NEBNext^® Globin & rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E7750 or NEB #E7755)
    or NEBNext^® rRNA Depletion Kit (Bacteria) (NEB #E7850 or NEB #E7860) 
    or NEBNext^® RNA Depletion Core Reagent Set (NEB #E7865 or NEB #E7870)
    or NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310 or NEB #E6350)

操作说明、说明书 & 用法

• 操作说明

  1. Where can I find protocols for using NEBNext^® RNA Library Prep reagents?

• 说明书

  产品说明书包含产品使用的详细信息、产品配方和质控分析。

  • manualE7760_E7765

• 应用实例

  • TechNote_NEBNext_Ultra_II_Directional_rRNA_Depletion
  • Obtain superior NGS library performance with lower input amounts using the NEBNext^® Ultra™ II Directional RNA Library Prep Kit for Illumina^®

FAQs & 问题解决指南

• FAQs

  1. I have USER enzyme from my library prep kit blue cap and USER enzyme from my NEBNext Index Primer kit red cap Are they the same
  2. What is the difference between the NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760) and the NEBNext Ultra II RNA Library Prep Kit (NEB #E7770)?
  3. What is the difference between the NEBNext Ultra II Directional RNA Library Prep Kit (NEB #E7760) and the original NEBNext Ultra Directional RNA Library Prep Kit (NEB #E7420)?
  4. What is the starting material I need to use when preparing libraries using the NEBNext Ultra II Directional RNA kit?
  5. Which kit can I use to isolate Poly (A) mRNA from Total RNA?
  6. Which kit can I use to deplete ribosomal RNA (rRNA) from human, mouse or rat RNA?
  7. Does the kit include adaptor and primers?
  8. Is it possible to reduce the volume of beads added during the second strand synthesis cleanup step of the protocol for E7760, E7765, E7770, and E7775?
  9. Which NEBNext Oligos can be used with this library prep kit?
  10. Can I use this NEBNext kit with adaptors and primers from other vendors than NEB?
  11. What do I do if I see a precipitate in the Ultra II End Prep Reaction Buffer?

• 问题解决指南

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