Next® 甲基化建库酶学转化法模块 |NEB酶试剂 New England Biolabs

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产品信息

下载技术说明

NEBNext 甲基化建库酶学转化法模块提供了一种高性能的酶学方法，能够代替重亚硫酸盐转化法来研究 DNA 甲基化。

在 EM-seq 实验流程中，由 TET2 氧化 5-mC 和 5-hmC 提供保护，避免在后续步骤中被 APOBEC 脱氨基。相反，未修饰的胞嘧啶被脱氨基成尿嘧啶。

特性：

  • 转化效率高
  • DNA 损伤极少
  • GC 偏嗜性极小
  • 5-mC 和 5-hmC 检测灵敏
  • 试剂盒同时包含用于文库制备的完整试剂（NEB #E7120）
图 1：NEBNext 酶学转化法甲基化建库和重亚硫酸盐转化方法

Next® 甲基化建库酶学转化法模块 |

产品类别:
Automation for NEBNext® NGS Library Prep Products,
Epigenetic Analysis Products,
Epigenetics Products

应用:
DNA Methylation Analysis,
Bisulfite Conversion,
Methylated DNA Analysis Products ,

Epigenetics

  • 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
    • E7125S     -20    
        Control DNA CpG methylated pUC19 E7122AVIAL -20 1 x 0.024 ml 0.1 ng/µl
        Control DNA CpG unmethylated Lambda E7123AVIAL -20 1 x 0.024 ml 2 ng/µl
        Elution Buffer E7124AVIAL -20 1 x 2.1 ml Not Applicable
        TET2 Reaction Buffer E7126AVIAL -20 1 x 0.3 ml Not Applicable
        TET2 Reaction Buffer Supplement E7127AVIAL -20 3 x 100 µl 100 X
        Oxidation Supplement E7128AVIAL -20 1 x 0.024 ml Not Applicable
        Oxidation Enhancer E7129AVIAL -20 1 x 0.024 ml Not Applicable
        TET2 E7130AVIAL -20 1 x 0.096 ml Not Applicable
        Fe (II) Solution E7131AVIAL -20 1 x 0.024 ml Not Applicable
        Stop Reagent E7132AVIAL -20 1 x 0.024 ml Not Applicable
        APOBEC E7133AVIAL -20 1 x 0.024 ml Not Applicable
        APOBEC Reaction Buffer E7134AVIAL -20 1 x 0.24 ml Not Applicable
        BSA E7135AVIAL -20 1 x 0.024 ml Not Applicable
        DTT E7139AAVIAL -20 1 x 0.5 ml 100 mM
    • E7125L     -20    
        Control DNA CpG methylated pUC19 E7122AAVIAL -20 1 x 0.096 ml 0.1 ng/µl
        Control DNA CpG unmethylated Lambda E7123AAVIAL -20 1 x 0.096 ml 2 ng/µl
        Elution Buffer E7124AAVIAL -20 1 x 8.6 ml Not Applicable
        TET2 Reaction Buffer E7126AAVIAL -20 1 x 1.2 ml Not Applicable
        TET2 Reaction Buffer Supplement E7127AAVIAL -20 3 x 400 µl 100 X
        Oxidation Supplement E7128AAVIAL -20 1 x 0.096 ml Not Applicable
        Oxidation Enhancer E7129AAVIAL -20 1 x 0.096 ml Not Applicable
        TET2 E7130AAVIAL -20 1 x 0.384 ml Not Applicable
        Fe (II) Solution E7131AAVIAL -20 1 x 0.096 ml Not Applicable
        Stop Reagent E7132AAVIAL -20 1 x 0.096 ml Not Applicable
        APOBEC E7133AAVIAL -20 1 x 0.096 ml Not Applicable
        APOBEC Reaction Buffer E7134AAVIAL -20 1 x 0.96 ml Not Applicable
        BSA E7135AAVIAL -20 1 x 0.096 ml Not Applicable
        DTT E7139AAVIAL -20 1 x 0.5 ml 100 mM

  • 特性和用法

    需要但不提供的材料

    • PCR 联管
    • 甲酰胺（Sigma #F9037-100 ml）或 0.1 N NaOH
    • 80% 乙醇
    • 0.1X TE，pH 8.0
    • 无核酶水
    • 纯化磁珠：NEBNext 纯化磁珠（随酶学转化法甲基化建库试剂盒提供，NEB #E7120）、SPRIselect® 试剂盒（Beckman Coulter, #B23317）、AMPure® XP 磁珠（Beckman Coulter, #A63881），或选择其它的磁珠制造商生产的磁珠产品。
    • 磁力架/支架，例如 NEBNext 磁性分离架（NEB #S1515）
    • PCR 仪
    • Bioanalyzer®、TapeStation® 和相关耗材或其它片段化分析仪

  • 注意事项

    1. FE(II) 溶液可能呈黄色。检测证明，颜色与性能无直接关系。

操作说明、说明书 & 用法

  • 操作说明

    1. Where can I find protocols for using the NEBNext® Enzymatic Methyl-seq Kit? (NEB #E7120, E7125, E7140)

  • 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。

    • manualE7125

FAQs & 问题解决指南

  • FAQs

    1. What types of sample can be processed using the NEBNext® Enzymatic Methyl-seq Kit and the Enzymatic Methyl-seq Conversion Module?
    2. What are the recommended inputs for the NEBNext® Enzymatic Methyl-seq Kit and the Enzymatic Methyl-seq Conversion Module?
    3. What is the difference between the NEBNext Enzymatic Methyl-seq Kit and the Enzymatic Methyl-seq Conversion Module?
    4. What buffers are recommended for shearing DNA in NEB’s enzymatic methyl-seq (EM-seq™) workflows?
    5. Can the “active” TET2 Buffer be stored longer than 4 months?
    6. Can the freshly diluted Fe(II) Solution be stored long term?
    7. Why, at some stages of the EM-seq protocol, do the NEBNext Sample Purification Beads behave differently when cleaning up the sample?
    8. Can enzymatically fragmented DNA be used as input material for EM-seq™?
    9. Can I use NaOH (sodium hydroxide) instead of formamide to denature my DNA prior to the deamination reaction?
    10. Is it normal if the FE(II) solution from the EM-seq product is yellow or a color change is observed?