上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。
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 1 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 #E7580 和 NEB #E7560 中包含用于多样本建库的、不同的 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
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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 -
E7300S -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 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 Nuclease-free Water E7327AVIAL -20 1 x 5 ml Not Applicable TE Buffer E7326AVIAL -20 1 x 0.48 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 6 Primer for Illumina E7316AVIAL -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 Gel Loading Dye, Blue E6138AVIAL -20 1 x 0.2 ml 6 X 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
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E7300L -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 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 Nuclease-free Water E7327AAVIAL -20 1 x 20 ml Not Applicable TE Buffer E7326AAVIAL -20 1 x 1.92 ml Not Applicable NEBNext Index 1 Primer for Illumina E7311AAVIAL -20 1 x 0.04 ml 10 µM NEBNext Index 2 Primer for Illumina E7312AAVIAL -20 1 x 0.04 ml 10 µM NEBNext Index 3 Primer for Illumina E7313AAVIAL -20 1 x 0.04 ml 10 µM NEBNext Index 4 Primer for Illumina E7314AAVIAL -20 1 x 0.04 ml 10 µM NEBNext Index 5 Primer for Illumina E7315AAVIAL -20 1 x 0.04 ml 10 µM NEBNext Index 6 Primer for Illumina E7316AAVIAL -20 1 x 0.04 ml 10 µM NEBNext Index 7 Primer for Illumina E7317AAVIAL -20 1 x 0.04 ml 10 µM NEBNext Index 8 Primer for Illumina E7318AAVIAL -20 1 x 0.04 ml 10 µM NEBNext Index 9 Primer for Illumina E7319AAVIAL -20 1 x 0.04 ml 10 µM NEBNext Index 10 Primer for Illumina E7320AAVIAL -20 1 x 0.04 ml 10 µM NEBNext Index 11 Primer for Illumina E7321AAVIAL -20 1 x 0.04 ml 10 µM NEBNext Index 12 Primer for Illumina E7322AAVIAL -20 1 x 0.04 ml 10 µM Gel Loading Dye, Blue E6138AAVIAL -20 1 x 1 ml 6 X 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
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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.） -
注意事项
- 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。
- 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。
操作说明、说明书 & 用法
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操作说明
- 6A. Quality Control Check and Size Selection using 6% Polyacrylamide Gel NEBNext Small RNA Library Prep Set for Illumina (E7300, E7580, E7560, E7330)
- 6B. Quality Control Check and Size Selection using Pippin Prep NEBNext Small RNA Library Prep Set for Illumina (E7300, E7580, E7560, E7330)
- 6C. Quality Control Check and Size Selection using AMPure XP Beads NEBNext Small RNA Library Prep Set for Illumina (E7300, E7580, E7560, E7330)
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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
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应用实例
- Automated size selection of NEBNext® Small RNA libraries with the Sage Pippin Prep™
FAQs & 问题解决指南
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FAQs
- Is it possible to prevent abundant, unwanted RNAs to be included in the Small RNA library?
- Can I use Total RNA to make small RNA libraries or do I have to isolate or enrich the sample for small RNA?
- Why does the RT primer hybridization occur before 5’adaptor ligation?
- Do I have to hybridize the RT primer again after 5’ ligation?
- How are the barcodes introduced in the Multiplex libraries?
- During size selection on 6% PAGE gel, which bands should I cut out of the gel?
- Are libraries prepared with the NEBNext® Small RNA Library prep kit for Illumina compatible with paired-end flow cells for cluster generation?
- Can I use the small RNA sample preparation kit for Directional-RNA sequencing?
- What is the sequence of the final PCR product?
- Can I use enriched small RNA instead of total RNA for the NEBNext Small RNA Library Prep for Illumina?
- What should I do if my Small RNA may not have 5´ Phosphate and 3´ OH groups?
- How should my NEBNext Small RNA library be trimmed?
- Are the Monarch RNA Cleanup Kits (NEB #T2030, #T2040, #T2050) compatible with NEBNext reagents for RNA library prep?
- What should I do if the sample sheet from the NEB.com website is not compatible with the software version I am using?
- Do you have Sample Sheets for use with the NEBNext Multiplex Small RNA Library Prep for Illumina?
- Are NEBNext adaptor and primers for Illumina compatible with NEBNext reagents for Illumina library preparation?
