Quick Blunting™ Kit |NEB酶试剂 New England Biolabs

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Quick Blunting™ Kit

The Quick Blunting™ Kit is used to convert DNA with incompatible 5´or 3´overhangs to 5´phosphorylated, blunt-ended DNA for efficient blunt-end ligation into DNA cloning vectors.

  • Restriction enzyme digested DNA is blunted in less than 30 minutes
  • Reactions are performed at room temperature in a ready-to-use mix
  • Suitable for restriction enzyme digested DNA, sheared or nebulized DNA or PCR product

精选视频

Quick Blunting™ Kit |

DNA Blunting Tutorial

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库存
货号 浓度 规格 目录价 北京 上海 广州 成都 苏州
E1201S 不适用 20 reactions ¥969.00 有 有 无 有 有
E1201L 不适用 100 reactions ¥3,909.00 有 无 无 无 无
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产品信息

The Quick Blunting Kit is used to convert DNA with incompatible 5´ or 3´ overhangs to 5´ phosphorylated, blunt-ended DNA for efficient blunt-end ligation into DNA cloning vectors. DNA is blunted using T4 DNA Polymerase (NEB #M0203) which has both 3´ → 5´ exonuclease activity and 5´ → 3´ polymerase activity. T4 Polynucleotide Kinase (NEB #M0201) is included in the enzyme mix for phosphorylation of the 5´ ends of blunt-ended DNA for subsequent ligation into a cloning vector. This kit is optimized for blunting up to 5 µg of DNA in a single reaction. To learn more about how to identify what type of overhang you have, visit this video tutorial.

Blunt Enzyme Mix supplied in: 100 mM KCl, 10 mM Tris-HCl(pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 0.1% Triton X-100 and 50% Glycerol.

1X Blunting Buffer:
100 mM Tris-HCl
50 mM NaCl
10 mM MgCl2
0.025% Triton X-100
5 mM dithiothreitol
pH 7.5 at 25°C

产品类别:
DNA Manipulation Products

应用:
Blunting,
Phosphorylation (Kinase),
PCR,

Fast Cloning: Accelerate your cloning workflows with reagents from NEB

  • 试剂盒组成

    Kit 组分

    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
    • E1201S     -20    
        Blunting Enzyme Mix M1201AVIAL -20 1 x 0.025 ml 不适用
        10X Blunting Buffer B1201SVIAL -20 1 x 0.5 ml 10 X
        Deoxynucleotide Solution Mix (1 mM) N1201AVIAL -20 1 x 0.1 ml 1 mM
    • E1201L     -20    
        Blunting Enzyme Mix M1201AAVIAL -20 1 x 0.125 ml 不适用
        10X Blunting Buffer B1201SVIAL -20 1 x 0.5 ml 10 X
        Deoxynucleotide Solution Mix (1 mM) N1201AAVIAL -20 1 x 0.5 ml 1 mM

  • 特性和用法

  • 优势和特性

    应用特性

    Prepare sheared, nebulized or restriction enzyme digested DNA for blunt-ended ligation into a plasmid, cosmid, fosmid or BAC vector

    Prepare PCR products for efficient blunt-end cloning

  • 相关产品

    相关产品

    • Quick Blunting™ and Quick Ligation™ Kits
    • Quick T4 DNA Ligase
    • Monarch® Plasmid Miniprep Kit 
    • Monarch® DNA Gel Extraction Kit
    • Monarch® PCR & DNA Cleanup Kit (5 μg)

  • 注意事项
    1. Restriction enzyme digested DNA can be blunted directly without purification. The Blunt Enzyme Mix has been optimized in Blunting Buffer, but is also active in NEBuffers 1,2,3 and 4, as well as BamHI, EcoRI and DpnII unique buffers when supplemented with dNTPs and dithiothreitol. There is a small reduction in ligation fidelity in these buffers. Transformation efficiency is lowest in NEBuffer 1 where the total yield is about 50% of optimum.
    2. ATP is not necessary for T4 Polynucleotide Kinase activity in the kit. The dATP and dTTP in the dNTP mix act as phosphate donors.
    3. The blunted reaction must be purified prior to phosphatase treatment by using a commercial purification kit, phenol extraction/ethanol precipitation, or gel electrophoresis.
    4. PCR generated DNA must be purified before blunting by using a commercial purification kit, such as Monarch® PCR & DNA Cleanup Kit (NEB #T1030), phenol extraction/ethanol precipitation, or gel electrophoresis.

操作说明、说明书 & 用法

  • 操作说明
    1. Blunting protocol for NEB PCR Cloning Kit
    2. Protocol for the Quick Blunting Kit (E1201)
    3. Transformation Protocol
    4. Quick Ligation Protocol (M2200)

工具 & 资源

  • 选择指南
    • Blunting Selection Chart

  • Web 工具
    • NEBcloner®

FAQs & 问题解决指南

  • FAQs
    1. Which ligase is recommended to ligate DNA treated with the Quick Blunting Kit?
    2. Can I use regular ligase to ligate my blunted product?
    3. I’ve blunted my DNA and now need to dephosphorylate the reaction with Antarctic Phosphatase. Do I need to purify the reaction?
    4. Does the PCR product need to be purified before blunting with the Quick Blunting Kit? What methods can be used to purify the product?
    5. I’ve digested my DNA and now want to blunt directly without purifying, can I add the blunting reagents directly to the restriction digest?
    6. I’ve blunted my sonicated gDNA for 15 minutes instead of the recommended 30 minute incubation time, will the reaction still work?
    7. I’ve accidentally skipped the heat-kill step after the blunting reaction. Will the ligation still work?

  • 问题解决指南
    • EpiMark® Methylated DNA Enrichment Kit Troubleshooting Guide

引用 & 技术文献

  • 引用文献

    产品引用文献查找工具

    Quick Blunting™ Kit | Powered by Bioz See more details on Bioz

质控、安全 & 法规

  • 质控分析
    每一新批次的 NEB 产品都要经过质控，以满足指定的质量标准，对特定产品的质量标准和每一批次的检测数据可以通过产品说明表格、COA、产品信息卡或者产品手册进行查阅或下载。关于 NEB 产品质控的详细信息，可从此处查阅。

  • 产品说明与变更通知

    产品说明与变更通知

    产品说明表包含产品的储存温度、有效期和规格，这些文件的命名规则如下：[货号]_[规格]_[版本]

    • E1201S_L_v1

  • CoA
    CoA 文件包含单个批次产品的储存温度、有效期和质控，这些文件的命名规则如下： [货号]_[规格]_[版本]_[Lot#]

    • E1201S_L_v1_0101801
    • E1201L_v1_10013484
    • E1201L_v1_10013485
    • E1201L_v1_10038381
    • E1201S_v1_10038380
    • E1201S_v1_10053797
    • E1201S_v1_10058037
    • E1201S_v1_10062856
    • E1201L_v1_10053799
    • E1201L_v1_10062855
    • E1201S_v1_10080108
    • E1201L_v1_10085502
    • E1201L_v1_10102455
    • E1201S_v1_10107427
    • E1201S_v1_10142224
    • E1201L_v1_10149861
    • E1201S_v1_10163391

  • SDS
    以下 SDS 文件可以帮助您安全地使用该产品

    • Blunting Enzyme Mix
    • 10X Blunting Buffer
    • Deoxynucleotide Solution Mix (1 mM)