Next Small RNA 多样本文库制备试剂盒（48 种 Index 引物） |NEB酶试剂 New England Biolabs

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产品信息

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 转换成含 Index 引物的文库，以便在 Illumina 平台上进行二次测序。新流程经优化，可最大限度减少接头二聚体的形成，同时产生产量高、多样性丰富的文库。

每个试剂盒组分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 引物的 small RNA 文库并在 Illumina 平台测序验证其功能。

 

NEBNext Small RNA 多样本文库制备试剂盒（48 种 Index 引物）

Next Small RNA 多样本文库制备试剂盒（48 种 Index 引物） |
产品类别:
Small RNA Library Preparation Products,
Next Generation Sequencing Library Preparation Products

  • 产品组分信息

    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
    • E7560S     -20    
        Gel Loading Dye, Blue E6138AAVIAL -20 1 x 1 ml 6 X
        NEBNext 3´ Ligation Enzyme Mix E7288AAVIAL -20 1 x 0.288 ml Not Applicable
        NEBNext Index 1 Primer for Illumina E7311AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 2 Primer for Illumina E7312AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 3 Primer for Illumina E7313AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 4 Primer for Illumina E7314AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 5 Primer for Illumina E7315AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 7 Primer for Illumina E7317AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 8 Primer for Illumina E7318AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 9 Primer for Illumina E7319AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 10 Primer for Illumina E7320AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 11 Primer for Illumina E7321AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 12 Primer for Illumina E7322AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 6 Primer for Illumina E7316AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext® Index 25 Primer for Illumina® E7561AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext® Index 27 Primer for Illumina® E7562AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext 3´ Ligation Reaction Buffer E7301AAVIAL -20 1 x 0.96 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
        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
        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 5´ SR Adaptor for Illumina E7328AVIAL -20 1 x 1,350 pmol Not Applicable
        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 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 Index 26 Primer for Illumina E7713AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 28 Primer for Illumina E7714AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 29 Primer for Illumina E7715AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 30 Primer for Illumina E7716AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 31 Primer for Illumina E7717AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 32 Primer for Illumina E7718AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 33 Primer for Illumina E7719AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 34 Primer for Illumina E7720AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 35 Primer for Illumina E7721AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 36 Primer for Illumina E7722AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 37 Primer for Illumina E7731AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 38 Primer for Illumina E7732AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 39 Primer for Illumina E7733AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 40 Primer for Illumina E7734AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 41 Primer for Illumina E7735AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 42 Primer for Illumina E7736AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 43 Primer for Illumina E7737AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 44 Primer for Illumina E7738AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 45 Primer for Illumina E7739AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 46 Primer for Illumina E7740AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 47 Primer for Illumina E7741AVIAL -20 1 x 0.01 ml 10 µM
        NEBNext Index 48 Primer for Illumina E7742AVIAL -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 24 Primer for Illumina E7592AVIAL -20 1 x 0.01 ml 10 µM
        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 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 18 Primer for Illumina E7586AVIAL -20 1 x 0.01 ml 10 µM
        Quick-Load pBR322 DNA-MspI Digest E7323AAVIAL -20 1 x 0.96 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® 离心过滤套管（醋酸纤维素过滤管）（Sigma Aldrich，#CLS8162）
    磁力架或磁力板（例如 NEBNext® Magnetic Separation Rack 磁性分离架，NEB #S1515S；Alpaqua ® 96S Super Magnet Plate 磁性分离板，#A001322；或同等物品）

    片段筛选所需材料：
        通过凝胶进行片段筛选：
            6% Novex® TBE PAGE 凝胶 1.0 mM
            10 孔板（Life Technologies 公司，#EC6265BOX）
            SYBR® Gold 核酸凝胶染料（Life Technologies 公司，#S-11494）
            RNase-free Disposable Pellet Pestles®（Kimble Kontes Asset Management 公司
            #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 公司，#A63881）

        通过 Pippin Prep™ 进行片段筛选：
            3% 不含染料的琼脂糖凝胶（Sage Science，#CDF 3010）

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

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

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?