Next® Small RNA 单样本文库制备试剂盒（1 种 Index 引物，与多样本兼容） |NEB酶试剂 New England Biolabs

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产品信息

Next® Small RNA 单样本文库制备试剂盒（1 种 Index 引物，与多样本兼容） |

NEBNext Small RNA 多样本文库制备试剂盒（48 种 Index 引物）包含接头、引物、酶和缓冲液，可将 small RNA 构建成含 Index 引物的文库，然后通过 Illumina 平台进行二代测序。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，可最大限度减少接头二聚体的形成。
 
请注意，NEBNext Small RNA 单样本文库制备试剂盒（1 种 Index 引物，与多样本兼容）可与您自己的 barcode 系统一起使用。此试剂盒包含接头，但不含多样本引物。如需 Index 引物，建议订购 NEB #E7300、NEB #E7580 或 NEB #E7560 试剂盒。
 
试剂盒内每个组份都通过严格的质量监控，每套试剂都经过 Small RNA 文库制备，并通过 Illumina 测序平台测序验证。
 

适用于 Illumina 测序平台的 Small RNA 文库制备流程
Next® Small RNA 单样本文库制备试剂盒（1 种 Index 引物，与多样本兼容） |

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

  • 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
    • E7330S     -20    
        NEBNext 3´ Ligation Reaction Buffer E7301AVIAL -20 1 x 0.24 ml Not Applicable
        Murine RNase Inhibitor E7308AVIAL -20 1 x 0.024 ml 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
        LongAmp™ Taq 2X Master Mix E7309AVIAL -20 1 x 1.2 ml Not Applicable
        Gel Loading Dye, Blue E6138AVIAL -20 1 x 0.2 ml 6 X
        Linear Acrylamide E7325AVIAL -20 1 x 0.048 ml Not Applicable
        Nuclease-free Water E7327AVIAL -20 1 x 5 ml Not Applicable
        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
        TE Buffer E7326AVIAL -20 1 x 0.48 ml Not Applicable
        NEBNext Index 1 Primer for Illumina E7329AVIAL -20 1 x 0.12 ml 10 µM
        NEBNext 3´ SR Adaptor for Illumina E7332AVIAL -20 1 x 0.024 ml Not Applicable
        NEBNext SR RT Primer for Illumina E7333AVIAL -20 1 x 0.024 ml Not Applicable
        NEBNext SR Primer for Illumina E7310AVIAL -20 1 x 0.06 ml Not Applicable
        NEBNext 5´ SR Adaptor for Illumina E7328AVIAL -20 1 x 1,350 pmol 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
        NEBNext 3´ Ligation Enzyme Mix E7288AVIAL -20 1 x 0.072 ml Not Applicable
    • E7330L     -20    
        NEBNext 3´ Ligation Reaction Buffer E7301AAVIAL -20 1 x 0.96 ml Not Applicable
        Murine RNase Inhibitor E7308AAVIAL -20 1 x 0.096 ml 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
        LongAmp™ Taq 2X Master Mix E7309AAVIAL -20 4 x 1.2 ml Not Applicable
        Gel Loading Dye, Blue E6138AAVIAL -20 1 x 1 ml 6 X
        Linear Acrylamide E7325AAVIAL -20 1 x 0.192 ml Not Applicable
        Nuclease-free Water E7327AAVIAL -20 1 x 20 ml Not Applicable
        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
        TE Buffer E7326AAVIAL -20 1 x 1.92 ml Not Applicable
        NEBNext Index 1 Primer for Illumina E7329AAVIAL -20 1 x 0.48 ml 10 µM
        NEBNext 3´ SR Adaptor for Illumina E7332AAVIAL -20 1 x 0.096 ml Not Applicable
        NEBNext SR RT Primer for Illumina E7333AAVIAL -20 1 x 0.096 ml Not Applicable
        NEBNext SR Primer for Illumina E7310AAVIAL -20 1 x 0.24 ml Not Applicable
        NEBNext 5´ SR Adaptor for Illumina E7328AVIAL -20 1 x 1,350 pmol 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
        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）

    QIAquick® PCR 纯化试剂盒（Qiagen®，#28104）
    Bioanalyzer®（Agilent Technologies, Inc.）

  • 注意事项

    1. 试剂盒优化

      试剂盒组份通过 FirstChoice® 人脑参考 RNA（Life Technologies, Inc. #AM7962）进行了整体优化。

      应使用高质量的总 RNA 作为起始样本。推荐使用 RNA 完整值（RIN）> 7 的样本。

      使用配备 Agilent RNA 6000 Nano 芯片的 Agilent 2100 Bioanalyzer 评估样本的质量和数量。

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. Are the Monarch RNA Cleanup Kits (NEB #T2030, #T2040, #T2050) compatible with NEBNext reagents for RNA library prep?
    14. How can I design my own i7 indexing/barcoding primer for additional barcoding options?