产品信息

NEBNext Small RNA 建库流程经过创新性优化，在保证制备高产量和多样性文库的同时，可以最大限度减少接头二聚体的形成。NEBNext Small RNA 文库制备试剂盒 2 包含接头和具有 12 种 Index 的多样本引物，可实现多样本建库。试剂盒 NEB #E7300 和 NEB #E7560 中包含用于多样本建库的额外 Index 引物。

每个试剂盒组份都必须通过严格的质量控制标准，每一套试剂都通过构建含 Index 的 small RNA 文库并在 Illumina 平台测序验证其功能。

GenomeWeb 的文章报道了 NEBNext Small RNA 试剂盒。

批次质控

所提供的单独批次均通过额外功能验证。单个试剂通过标准酶活性和质控分析，并同时满足各单独组分页面列出的严格的附加质控标准

产品类别:

Small RNA Library Preparation Products,

Next Generation Sequencing Library Preparation Products

• 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
  E7580S     -20       NEBNext 3´ Ligation Reaction Buffer E7301AVIAL -20 1 x 0.24 ml Not Applicable   NEBNext 3´ SR Adaptor for Illumina E7332AVIAL -20 1 x 0.024 ml Not Applicable   NEBNext 5´ SR Adaptor for Illumina E7328AVIAL -20 1 x 1,350 pmol Not Applicable   NEBNext 5´ Ligation Reaction Buffer E7304AVIAL -20 1 x 0.024 ml Not Applicable   NEBNext 5´ Ligation Enzyme Mix E7305AVIAL -20 1 x 0.06 ml Not Applicable   NEBNext SR RT Primer for Illumina E7333AVIAL -20 1 x 0.024 ml Not Applicable   NEBNext First Strand Synthesis Reaction Buffer E7334AVIAL -20 1 x 0.192 ml Not Applicable   ProtoScript II Reverse Transcriptase E7355AVIAL -20 1 x 0.024 ml Not Applicable   Murine RNase Inhibitor E7308AVIAL -20 1 x 0.024 ml Not Applicable   LongAmp™ Taq 2X Master Mix E7309AVIAL -20 1 x 1.2 ml Not Applicable   NEBNext SR Primer for Illumina E7310AVIAL -20 1 x 0.06 ml Not Applicable   Gel Loading Dye, Blue E6138AVIAL -20 1 x 0.2 ml 6 X   Quick-Load pBR322 DNA-MspI Digest E7323AVIAL -20 1 x 0.24 ml Not Applicable   DNA Gel Elution Buffer E7324AVIAL -20 1 x 12 ml Not Applicable   Linear Acrylamide E7325AVIAL -20 1 x 0.048 ml Not Applicable   TE Buffer E7326AVIAL -20 1 x 0.48 ml Not Applicable   Nuclease-free Water E7327AVIAL -20 1 x 5 ml Not Applicable   NEBNext Index 13 Primer for Illumina E7581AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 14 Primer for Illumina E7582AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 15 Primer for Illumina E7583AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 16 Primer for Illumina E7584AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 17 Primer for Illumina E7585AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 18 Primer for Illumina E7586AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 19 Primer for Illumina E7587AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 20 Primer for Illumina E7588AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 21 Primer for Illumina E7589AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 22 Primer for Illumina E7590AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 23 Primer for Illumina E7591AVIAL -20 1 x 0.01 ml 10 µM   NEBNext Index 24 Primer for Illumina E7592AVIAL -20 1 x 0.01 ml 10 µM   NEBNext 3´ Ligation Enzyme Mix E7288AVIAL -20 1 x 0.072 ml Not Applicable
  E7580L     -20       NEBNext 3´ Ligation Reaction Buffer E7301AAVIAL -20 1 x 0.96 ml Not Applicable   NEBNext 3´ SR Adaptor for Illumina E7332AAVIAL -20 1 x 0.096 ml Not Applicable   NEBNext 5´ SR Adaptor for Illumina E7328AVIAL -20 1 x 1,350 pmol Not Applicable   NEBNext 5´ Ligation Reaction Buffer E7304AAVIAL -20 1 x 0.096 ml Not Applicable   NEBNext 5´ Ligation Enzyme Mix E7305AAVIAL -20 1 x 0.24 ml Not Applicable   NEBNext SR RT Primer for Illumina E7333AAVIAL -20 1 x 0.096 ml Not Applicable   NEBNext First Strand Synthesis Reaction Buffer E7334AAVIAL -20 1 x 0.768 ml Not Applicable   ProtoScript II Reverse Transcriptase E7355AAVIAL -20 1 x 0.096 ml Not Applicable   Murine RNase Inhibitor E7308AAVIAL -20 1 x 0.096 ml Not Applicable   LongAmp™ Taq 2X Master Mix E7309AAVIAL -20 4 x 1.2 ml Not Applicable   NEBNext SR Primer for Illumina E7310AAVIAL -20 1 x 0.24 ml Not Applicable   Gel Loading Dye, Blue E6138AAVIAL -20 1 x 1 ml 6 X   Quick-Load pBR322 DNA-MspI Digest E7323AAVIAL -20 1 x 0.96 ml Not Applicable   DNA Gel Elution Buffer E7324AAVIAL -20 1 x 48 ml Not Applicable   Linear Acrylamide E7325AAVIAL -20 1 x 0.192 ml Not Applicable   TE Buffer E7326AAVIAL -20 1 x 1.92 ml Not Applicable   Nuclease-free Water E7327AAVIAL -20 1 x 20 ml Not Applicable   NEBNext Index 13 Primer for Illumina E7581AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 14 Primer for Illumina E7582AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 15 Primer for Illumina E7583AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 16 Primer for Illumina E7584AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 17 Primer for Illumina E7585AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 18 Primer for Illumina E7586AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 19 Primer for Illumina E7587AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 20 Primer for Illumina E7588AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 21 Primer for Illumina E7589AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 22 Primer for Illumina E7590AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 23 Primer for Illumina E7591AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext Index 24 Primer for Illumina E7592AAVIAL -20 1 x 0.04 ml 10 µM   NEBNext 3´ Ligation Enzyme Mix E7288AAVIAL -20 1 x 0.288 ml Not Applicable

• 特性和用法

  需要但不提供的材料

  3 M 乙酸钠，pH 5.5
  100% 乙醇
  80% 乙醇
  Corning^®、Costar^®、Spin-X^® 离心过滤管（醋酸纤维素过滤器）（Sigma Aldrich，#CLS8162）
  磁力架或磁力板（例如：NEBNext^® 磁性分离架，NEB #S1515S；Alpaqua ^® 96S 超磁性分离板，#A001322；或同等器材）

  片段筛选所需材料
  通过切胶进行片段筛选：
  6% Novex^® TBE PAGE gel，1.0 mM
  10 孔板（Life Technologies, Inc. #EC6265BOX）
  SYBR^® Gold 核酸凝胶染料（Life Technologies, Inc. #S-11494）
  RNase-free Disposable Pellet Pestles^®（Kimble Kontes Asset Management, Inc. #749521-1590）
  干冰/甲醇冰浴或 –80℃ 冰箱

  通过磁珠进行片段筛选：
  Agencourt^® AMPure^® XP Beads（Beckman Coulter, Inc. #A63881）

  通过 Pippin Prep™ 进行片段筛选：
  3% Agarose Dye Free Gel（Sage Science，#CDF 3010）

  Monarch^® PCR & DNA 纯化试剂盒（5 μg）（NEB# T1030）
  Bioanalyzer®^®（Agilent^® Technologies, Inc.）

操作说明、说明书 & 用法

• 操作说明

  1. 6A. Quality Control Check and Size Selection using 6% Polyacrylamide Gel  NEBNext Small RNA Library Prep Set for Illumina (E7300, E7580, E7560, E7330)
  2. 6B. Quality Control Check and Size Selection using Pippin Prep NEBNext Small RNA Library Prep Set for Illumina (E7300, E7580, E7560, E7330)
  3. 6C. Quality Control Check and Size Selection using AMPure XP Beads NEBNext Small RNA Library Prep Set for Illumina (E7300, E7580, E7560, E7330)

• 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。

  • manualE7300_E7330_E7560_E7580

• 应用实例

  • Automated size selection of NEBNext^® Small RNA libraries with the Sage Pippin Prep™

FAQs & 问题解决指南

• FAQs

  1.  Is it possible to prevent abundant, unwanted RNAs to be included in the Small RNA library?
  2. Can I use Total RNA to make small RNA libraries or do I have to isolate or enrich the sample for small RNA?
  3. Why does the RT primer hybridization occur before 5’adaptor ligation?
  4. Do I have to hybridize the RT primer again after 5’ ligation?
  5. How are the barcodes introduced in the Multiplex libraries?
  6. During size selection on 6% PAGE gel, which bands should I cut out of the gel?
  7. Are libraries prepared with the NEBNext^® Small RNA Library prep kit for Illumina compatible with paired-end flow cells for cluster generation?
  8. Can I use the small RNA sample preparation kit for Directional-RNA sequencing?
  9. What is the sequence of the final PCR product?
  10. Can I use enriched small RNA instead of total RNA for the NEBNext Small RNA Library Prep for Illumina?
  11. What should I do if my Small RNA may not have 5´ Phosphate and 3´ OH groups?
  12. How should my NEBNext Small RNA library be trimmed?
  13. What should I do if the sample sheet from the NEB.com website is not compatible with the software version I am using?
  14. Do you have Sample Sheets for use with the NEBNext Multiplex Small RNA Library Prep for Illumina?
  15. Are NEBNext adaptor and primers for Illumina compatible with NEBNext reagents for Illumina library preparation?