Next Ultra II RNA Library Prep with Sample Purification Beads |NEB酶试剂 New England Biolabs

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产品信息

Next Ultra II RNA Library Prep with Sample Purification Beads |
 

The Ultra II RNA Library Prep Kit for Illumina is designed for non-directional (non-strand-specific) RNA library construction, and 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) mRNA enrichment, the kit enables the production of high quality libraries from 10 ng of Total RNA, respectively, up to 1µg.

For directional (strand-specific) RNA library preparation, see the NEBNext Ultra II Directional RNA Library Prep kit for Illumina.

See what customers are saying about NEBNext Ultra II 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 (link to E7770 Complexity and Transcript Coverage daughter pages)
  • Optimize your time with streamlined workflows, reduced hands-on time, and automation compatibility
  • Rely on robust performance
  • Compatible with NEBNext poly(A) mRNA Isolation, rRNA Depletion reagents and multiplexing adaptors and primers

Also available without optional SPRIselect® beads.

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.

LIBRARY YIELDS

Figure 1. NEBNext Ultra II RNA produces the highest yields, from a range of input amounts
Next Ultra II RNA Library Prep with Sample Purification Beads |
Poly(A)-containing mRNA was isolated from Human Universal Reference RNA (Agilent #740000) and libraries were prepared using the NEBNext Ultra II RNA Kit (plus the NEBNext Poly(A) mRNA Magnetic Isolation Module), and the Illumina TruSeq RNA Sample Preparation Kit v2. Input RNA and number of PCR cycles are indicated. Library yields from an average of three replicates are shown.

View additional data on library yields

DUPLICATION RATES

Figure 2. NEBNext Ultra II RNA with NEBNext rRNA Depletion results in lower duplication rates .
Next Ultra II RNA Library Prep with Sample Purification Beads |
Ribosomal RNA was depleted from 1 μg, 100 ng and 5 ng of Human Universal Reference RNA (Agilent #740000) using the NEBNext rRNA Depletion Kit (Human/ Mouse/Rat) or Illumina Ribo-Zero™ Gold rRNA Removal Kit (Human/Mouse/Rat). Libraries were then prepared using the NEBNext Ultra II RNA Kit or the Illumina TruSeq RNA Library Prep Kit v2, respectively. 5 ng input was tested only with the NEBNext kits. Libraries were sequenced on an Illumina NextSeq® 500 using paired-end mode (2×76 bp). Reads were down sampled to 10 million read pairs and mapped to the hg19 reference genome. Duplication rates were calculated as a fraction of uniquely mapped reads using the ‘Read Duplication’ tool of RSeQC where reads mapping to the same genomic location are regarded as duplicated reads.

View additional data on library quality

MAXIMIZING TRANSCRIPT COVERAGE

Figure 3. NEBNext Ultra II RNA libraries provide uniform coverage across the gene body of transcripts
Next Ultra II RNA Library Prep with Sample Purification Beads |
Poly(A)-containing mRNA was isolated from Human Universal Reference RNA (Agilent #740000), and libraries were prepared using the NEBNext Ultra II RNA Kit (plus the NEBNext Poly(A) mRNA Magnetic Isolation Module), and the Illumina TruSeq RNA Sample Preparation Kit v2. Input RNA amount is indicated. Libraries were sequenced on an Illumina® NextSeq® 500 using paired-end mode (2×76 bp). This view of the 5´ to 3´ coverage of RefSeq transcripts reveals consistent coverage for Ultra II RNA libraries as input RNA is decreased from 1 μg to 10 ng. The changes apparent in The TruSeq kit results from loss of coverage at the 3´ end of some transcripts.

View additional data on transcript coverage

EXCELLENT LIBRARY COMPLEXITY AT LOW INPUT AMOUNTS

Figure 4. Low input NEBNext Ultra II RNA libraries retain complexity even at low input amounts
Next Ultra II RNA Library Prep with Sample Purification Beads |
Ribosomal RNA was depleted from 1 μg, 100 ng and 5 ng of Human Universal Reference RNA (Agilent #740000) with recommended amounts of ERCC RNA Spike-In Mix I (Thermo Fisher Scientific #4456740) using the NEBNext rRNA Depletion Kit (Human/Mouse/Rat) and libraries were then prepared using the NEBNext Ultra II RNA Kit. Libraries were sequenced on an Illumina NextSeq® 500 using paired-end mode (2×76 bp). Salmon 0.4.0 was used for read mapping and quantification of all ERCC transcripts. R2 values for linear fit are shown. TPM (Transcripts Per Kilobase Million) correlation analysis of the transcripts indicates excellent transcript expression correlation between the different inputs for Ultra II RNA libraries (A), including ERCC transcripts (B).

View additional data on library complexity

产品类别:
RNA Library Prep for Illumina

  • 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
    • E7775S     Multi-temperature    
        NEBNext® Ultra™ II RNA Library Prep Kit for Illumina® E7770S -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 E7427AVIAL -20 1 x 0.192 ml Not Applicable
        Nuclease-free Water E7764AVIAL -20 1 x 1.25 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
        NEBNext® Sample Purification Beads E7767S 25    
        NEBNext® Sample Purification Beads E7767SVIAL 25 1 x 6.65 ml Not Applicable
    • E7775L     Multi-temperature    
        NEBNext® Ultra™ II RNA Library Prep Kit for Illumina® E7770L -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 E7427AAVIAL -20 1 x 0.768 ml Not Applicable
        Nuclease-free Water E7764AAVIAL -20 1 x 5.7 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
        NEBNext® Sample Purification Beads E7767L 25    
        NEBNext® Sample Purification Beads E7767SVIAL 25 4 x 6.65 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)
    • 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 Ultra II Directional RNA Library Prep Kit for Illumina (NEB #E7770 & #E7775)?

  • 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。

    • manualE7770_E7775

FAQs & 问题解决指南

  • FAQs

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

  • 问题解决指南

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