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DNA甲基化极速修饰试剂盒
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p-1026-9

DNA甲基化极速修饰试剂盒

DNA甲基化极速修饰试剂盒,非常适合做MSP,实时定量荧光定量PCR即qMSP,MS-HRM,甲基化微阵列,及甲基化测序分析(焦磷酸测序和深度测度)等下游操作。更多视频请关注视频号【艾维缔】。哔哩哔哩【IVDSHOW】。抖音【军哥聊表观】。
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    胞嘧啶环中 5-碳上的甲基若具备共有原子价的条件,就 能引发DNA 的甲基化,产生5-甲基胞嘧啶。现在有多种方法 可用于DNA 甲基化状态的分析。但是,只有基因组DNA亚硫酸氢盐修饰,紧接着 PCR 扩增,克隆,以及个体 PCR 扩增引物排序,能对个体DNA 分子上的个体胞嘧啶的甲基化状态提供可靠的信息。通过使用亚硫酸氢盐处理 DNA,胞嘧啶 残留物将被脱氨基成尿嘧啶,而 5-甲基胞嘧啶则被完整的保留。

    传统的亚硫酸氢盐转化方法需要12-16个小时进行亚硫 酸氢盐处理,DNA严重降解(>80%), 高不相称的甲基胞嘧啶脱氨基(>3.5%),低胞嘧啶转化率<95%)。在2005年三月份,我司成为第一个研发快速DNA亚硫酸氢盐修饰方法的公司,我司的方法客服了其他方法所存在的一系列问题—— 将亚硫酸氢盐过程从16小时缩短至只需要1.5小时,显著的提高了胞嘧啶转化效率(>99.9%),并有效的防止了修饰后的DNA降解。
    为了有效的配备转化后的DNA用于各种下游分析,一种理想的亚硫酸氢盐修饰方法应该具备(1)高度精确允许胞嘧啶完全转化为尿嘧啶(正确的转化)而不会产生甲基胞嘧啶 脱氨基成为胸腺嘧啶(不正确的转化);(2)极速,使亚硫酸氢盐过程尽可能的缩短,因为对于基本研究,特别是临床应用,都要求要进行快速DNA甲基化分析。
我司再接再厉创新研发DNA甲基化极速修饰试剂盒,完美的演绎了DNA亚硫酸氢盐修饰方法,从而取得更好的DNA甲基化分析。
    该试剂盒极大的改进了目前所使用的DNA亚硫酸氢盐修饰试 剂盒。具备新颖且能最大利用的亚硫酸氢盐合成物,该DNA 修饰试剂盒能够使DNA修饰步骤缩短在20分钟之内,并获得胞嘧啶完全充分的转化结果。更重要的是,它能极大的减少5-甲基胞嘧啶不正确转化为胸胞嘧啶(<0.1%)。该DNA修饰试剂盒适用于MS-PCR,实时定量MS-PCR,以及甲基化微阵列。
胞嘧啶(C)完全转化:用DNA甲基化极速修饰试剂盒处理从 3 种癌细胞系中分离的 200 ng基因组 DNA。然后,使用未转化 DNA 特异性引物和已转化 DNA 特异性引物(β-actin,110 bps)分别测定每个处理样本中未转化 DNA 和已转化 DNA 的含量。A:实时 PCR;B:终点 PCR。DNA甲基化极速修饰试剂盒处理过的 DNA 已完全转化,45 个循环后,处理过的样品中没有未转化的 DNA。
有效保护 DNA: 使用 DNA甲基化极速修饰试剂盒转换不同数量(0.2 ng-200 ng)的完全甲基化人类基因组 DNA。1 µl 的 20 µl 洗脱液用于实时 qPCR,一对引物用于扩增转化后的 DNA。使用 DNA甲基化极速修饰试剂盒,只要 0.2 ng DNA 就足以进行亚硫酸氢盐转化。A:实时 PCR;B:起始 DNA 量-CT 值曲线。
        开发的DNA甲基化极速修饰试剂盒。是专门针对提取好的DNA而设计的,具有如下的优势和特性:
方便的单一温度孵育。不需要进行单独的DNA变性步骤。
最快速最方便的操作步骤,能在30分钟左右完成。
完全的将未甲基化的胞嘧啶转化为尿嘧啶(>99.99%), 不合适的和错误的转化甲基胞嘧啶转化为胸腺嘧啶极其 微小(<0.1%),可以忽略。
强劲保护DNA不被降解,能保证90%DNA不被损失掉。
只需要输入极其少量的DNA进行修饰即可——0.2 ng或 者 50个细胞。
简单,可靠,不变的修饰条件。

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型号:A-P-1026-050(50)

型号:A-P-1026-200(200)

储存温度

BF1

Conversion Mix Solution

6 ml 12 ml 室温
BF2 Capture Solution 15 ml 30 ml 室温
BF3 Desulphonation Solution 60 ul 120 ul 室温
BF4 Elution Solution 1 ml 2 ml 室温
BF5

Conversion Enhancer

5 瓶 20 瓶 室温
BF6 Denaturation Enhancer 600 ul 1.2 ml 室温
BF7 F-spin Column* 50个 200个 室温

BF8

F-Collection Tube

50个 200个 室温

操作手册

 

1

1

室温

*注意:在将离心柱放入微型离心机之前,始终将离心柱盖子旋紧盖严

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警告 本品仅供科研使用,请勿用于临床与诊断。

 

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