标签归档：conversion

EpiSight(TM) Bisulfite Conversion Kit Ver.2 EpiSight™亚硫酸氢盐转化试剂盒2

发表于由

EpiSight(TM) Bisulfite Conversion Kit Ver.2

EpiSight™亚硫酸氢盐转化试剂盒2

品牌：FUJIFILM Wako
CAS No.：
储存条件：2-10℃
纯度：–
产品编号

(生产商编号)

 等级 规格 运输包装 零售价(RMB) 库存情况 参考值

291-78501

 for Genetic Research 20Tests – 咨询 – –


* 干冰运输、大包装及大批量的产品需酌情添加运输费用


* 零售价、促销产品折扣、运输费用、库存情况、产品及包装规格可能因各种原因有所变动，恕不另行通知，确切详情请联系上海金畔生物科技有限公司。

EpiMark® Bisulfite Conversion Kit |NEB酶试剂 New England Biolabs

发表于由

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

Genomic DNA from many organisms has modified nucleotides. In the mammalian genome, the modified base is predominately 5-methylcytosine (5-mC), which is involved in gene expression regulation, including selective inactivation of one X chromosome in females of mammalian species (1,2). Symmetrical CpG DNA methylation is heritable, but also reversible. This change of methylation status results in an enormous number of combinations of epigenetic states that can regulate gene expression. In mammalian cells, DNA methylation mainly occurs in CpG dinucleotides and is carried out by two methyltransferase enzymatic activities, namely, maintenance methylation and de novo methylation. The maintenance methyltransferase, DNMT1, is involved in the DNA methylation after every cellular DNA replication cycle. DNMT3a and DNMT3b are the de novo methyltransferases that are active in early development.

For optimal results, 50 ng–2 μg DNA is recommended.

Kit Components: 
Each EpiMark Bisulfite Conversion Kit contains sufficient reagents to perform the bisulfite conversion reaction of 48 samples. Store at room temperature (15–20°C). However, for long-term storage (longer than 4 weeks) Desulphonation Reaction Buffer concentrate should be stored at 2–8°C. Prepared Bisulfite Mix can be stored at -20°C for up to six months. 

产品类别:
Discontinued (<3 years)

  • 特性和用法

    需要但不提供的材料

    Custom design primers
    EpiMark Hot Start Taq DNA Polymerase (NEB #M0490) or other hot start DNA polymerase specific for bisulfite converted DNA.
    Heat block or water bath (requiring temperatures of 65°C and 92°C)
    96–100% molecular biology grade ethanol
    PCR Thermal Cycler
    0.2 ml strip tubes and caps
    1.5 ml reaction tubes
    Nuclease-free Water
    Pipettors and pipette tips (for minimization of cross contamination, use aerosol
    barrier tips)

  • 方法概述

    方法概述

    Many tools can be used to determine the methylaton status of DNA, including antibodies, methyl binding proteins, methylation-dependent restriction enzymes, and most recently next generation sequencing. However, the most commonly used technique to date is sodium bisulfite conversion, the “gold standard” for methylation analysis (3). Incubation of the target DNA with sodium bisulfite results in conversion of all unmodified cytosines to uracils leaving the modified bases (5-mC or 5-hmC) intact. (see Figure 1). The most critical step in methylation analysis using bisulfite conversion is the complete conversion of unmodified cytosines. This is achieved by alternating cycles of thermal denaturation with incubation reactions. The protocol optimized for this kit is simple and gives consistent results.

    Figure 1. Overview of Bisulfite Conversion ReactionEpiMark® Bisulfite Conversion Kit |
    Methylated cytosines are protected and remain unchanged, while unmethylated cytosines are deaminated to uracil after treatment with sodium bisulfite.
    Figure 2. Performance evaluation of the EpiMark Bisulfite Conversion Kit.EpiMark® Bisulfite Conversion Kit |
    One μg of genomic DNA was bisulfite treated using the Epimark Bisulfite Conversion Kit, and 2 μl of eluted DNA was analyzed by end-point PCR. 388, 544, and 731 bp amplicons were amplified with primer pairs for bisulfite converted DNA (lanes 1, 3, and 5), or with primer pairs for unconverted DNA (lanes 2, 4, and 6). Epimark Hot Start Taq DNA Polymerase was used for amplification. Lanes 2, 4, and 6 clearly show no amplification product, indicating complete DNA conversion. Marker M is the 2-Log DNA Ladder (NEB #N3200).
    Figure 3. Conversion of a methylated DNA fragment.EpiMark® Bisulfite Conversion Kit |
    50 nanograms of methylated plasmid DNA was bisulfite converted and a 146 bp DNA fragment was amplified using Epimark Hot Start Taq DNA Polymerase (NEB #M0490). Amplicons were cloned, and individual clones were sequenced to determine the methylation status. Sequence aligment of 10 clones is shown to demonstrate conversion of unmethylated cytosine to thymine, while the unconverted methylated cytosine remains intact. The known methylated cytosines at the CpG site shown in red, converted cytosines marked as T, and rest of the sequence is shown as a dashed line.

  • 相关产品

    相关产品

    • EpiMark® 热启动 Taq DNA 聚合酶

  • 参考文献

    1. Bestor, T.H., Verdine, G.L. (1994). Curr. Opin. Cell Biol. . 6, 380-389.
    2. Ooi, S.K., O’Donnell A.H., Bestor T.H. (2009). J. Cell Sci. 122, 2787-2791.
    3. Frommer, M., et. al. (1992). Proc. Natl. Acad. Sci. USA. 89, 1827-1831.
    4. Kumaki, Y., Oda, M. and Okano, M. (2008). Nucleic Acids Res. . 36,

操作说明、说明书 & 用法

  • 操作说明

    1. Reagent Preparation Using EpiMark® Bisulfite Conversion Kit (E3318)
    2. End-point PCR Using EpiMark® Bisulfite Conversion Kit (E3318)
    3. Cycling Protocol Using EpiMark™ Bisulfite Conversion Kit (E3318)

  • 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。

    • manualE3318

工具 & 资源

  • 选择指南

    • DNA Methylation Table

FAQs & 问题解决指南

  • FAQs

    1. How should I store my bisulfite converted DNA after it is eluted from the column?
    2. Does my sample need to be free of RNA for bisulfite conversion?
    3. No PCR product is detected when I amplify more than 300 bp amplicon.
    4. Do I need extra purification of my mammalian DNA prep in order to use it in a bisulfite conversion reaction?
    5. Which DNA polymerase do you recommend for amplification of converted DNA?
    6. My thermocycler does not allow to set up the reaction volume to 140 μl, what I should do?
    7. What is the shelf life of the reagents supplied?

Conversion Adapter for TK-CP384

发表于由

Conversion Adapter for TK-CP384

品牌：Tokken
CAS No.：
储存条件：室温
纯度：–
产品编号

(生产商编号)

 等级 规格 运输包装 零售价(RMB) 库存情况 参考值

TK-CP96-384

 – 1 piece – 咨询 – –


* 干冰运输、大包装及大批量的产品需酌情添加运输费用


* 零售价、促销产品折扣、运输费用、库存情况、产品及包装规格可能因各种原因有所变动，恕不另行通知，确切详情请联系上海金畔生物科技有限公司。