·产品描述

   Methylamp全细胞亚硫酸氢盐修饰试剂盒是本公司的创新独有的技术，提供了一整套必需的试剂和方法，运用Epigentek独创的技术对亚硫酸氢盐法进行了简化和改进，来对DNA进行甲基化分析.整个过程耗时仅3小时，得到比别的试剂盒更好的分析结果.Methylamp全细胞亚硫酸氢盐修饰试剂盒专门用于针对初始原料（如96/384孔板上培养的细胞、组织切片样品、显微解剖样品、活组织切片已经早期胚胎细胞/卵母细胞）研究DNA甲基化.
   已修饰的DNA用本试剂盒洗脱后可直接用于MS-PCR，也广泛适用于所有DNA甲基化分析相关技术，其中包括常规MS-PCR、亚硫酸氢盐测序、焦磷酸测序以及甲基化芯片.如果产物用来做MSP， PCR的循环数最好大于45.每种修饰实验的初始材料的量可以在100-20000个细胞，或者1 ug-100 ug组织，抑或0.2-2 mm2组织切片样品的范围.为了达到最佳的修饰效果，宜把范围分别控制在500- 5000个细胞，或5-20 ug组织，抑或0.5-1mm2组织切片样品.

   Methylamp全细胞亚硫酸氢盐修饰试剂盒含有直接在细胞或组织中进行亚硫酸氢盐转换实验所需的所有试剂.本试剂盒用特殊的反应缓冲液能将DNA从细胞或组织中纯化出来，并在同一个管子将其热变性的同时进行亚硫酸氢盐转换修饰.在修饰处理过程中，亚硫酸氢盐试剂与单链DNA反应，将胞嘧啶转换成尿嘧啶.修饰缓冲液中含有本试剂盒特有的DNA预备反应物，可以专利亚硫酸氢盐处理样品过程DNA化学或热降解.无毒的已修饰DNA俘获缓冲液能将DNA牢牢的结合在柱子的滤膜上，可以放心地洗涤DNA上的亚硫酸氢钠和盐离子.被修饰的DNA随后可以被洗脱下来，储存在-20℃两个月以上.该全细胞亚硫酸盐DNA甲基化修饰试剂盒内主要产品组份包括：W1 （Digestion powder） W2 （Digestion solution） W3 （Cell collection buffer） W4 （DNA modification powder） W5 （DNA modification buffer） W6 （Balance buffer） W7 （DNA binding buffer） W8 （modified DNA elution） F-spin column F-collection tube 用户指南手册

·产品特点

1、直接修饰细胞或组织中的DNA； 

2、快速地流水线式的3小时操作过程； 

3、将未甲基化的胞嘧啶完全转换为尿嘧啶：修饰DNA占总量的99、5%以上； 

4、最少的DNA 降解量：利用一种本公司特有的DNA保护缓冲液，减少90%的DNA损失；

·产品说明书

The Methylamp™ Whole Cell Bisulfite Modification Kit is an innovative and unique set of essential components which enables the experimenter to perform DNA methylation analysis and modify DNA directly from cells or tissues using Epigentek's uniquely simplified and streamlined bisulfite method. The entire procedure can be completed within only 3 hours. The Methylamp™ Whole Cell Bisulfite Modification Kit is specifically designed for DNA methylation research using minute amounts of starting materials including cells cultured in 96-well/384-well plates, tissue section samples, microdissection samples, tissue biopsy and early embryonic cells/oocytes. 

Modified DNA is eluted by using the Methylamp™ Whole Cell Bisulfite Modification Kit and is suitable for real time MS-PCR. It is also suitable for all techniques currently used for the analysis of DNA methylation; including conventional MS-PCR, bisulfite sequencing, pyrosequencing, and methylation microarray. If you use the Methylamp™ Whole Cell Bisulfite Modification Kit for MSP, the numbers of PCR cycles should be greater than 45. The kit has the following advantages:

• Directly modify DNA in cells or tissues.
• Rapid streamlined 3 hour procedure.
• Completely converts unmethylated cytosine into uracil: modified DNA > 99.5%.
• The lowest degradation of DNA in the modification process: more than 90% of DNA loss can be prevented with a unique DNA protection buffer.

Principle & Procedure
The Methylamp™ Whole Cell Bisulfite Modification Kit contains all reagents required for bisulfite conversion directly on a cell or tissue sample. The kit allows DNA to be isolated from cells or tissues, denatured and bisulfite modified simultaneously in same tube with the specific reaction buffer under the thermodynamic condition. In the modification process, bisulfite reagent reacts specifically with single-stranded DNA, thereby deaminating cytosine and creating a uracil residue. The unique DNA protection reagents contained in the modification buffer can prevent the chemical and thermophilic degradation of DNA in the bisulfite treatment. The non-toxic modified DNA capture buffer enables DNA to tightly bind to the column filter, thus DNA cleaning can be carried out on the column to effectively remove residual sodium bisulfite and salts. Modified DNA can then be eluted and stably stored at -20°C for up to 2 months.

Starting Materials
The amount of starting materials for each modification can be 100-20000 cells, or 1 µg-100 µg of tissue, or 0.2-2 mm2 tissue section samples. For optimal modification, the amount should be 500-5000 cells, or 5-20 µg of tissue, or 0.5-1 mm2 tissue section samples, respectively.

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