·产品描述

Methylamp DNA修饰试剂盒包含DNA样本亚硫酸氢盐转换修饰所需要的全部试剂.DNA与亚硫酸氢盐试剂反应，能特异性的使DNA化学变性，产生单链DNA，从而使胞嘧啶脱氨基形成尿嘧啶.修饰液中含有的独特的DNA保护剂，能在亚硫酸氢盐处理后阻止化学性或者嗜热性降解DNA.无毒性的修饰DNA捕获液能使DNA紧密的绑定在圆柱过滤器，因此DNA的清洗能有效地去除残余的亚硫酸氢钠或者盐.然后修饰的DNA洗提，储存在-20度可以保持2个月.

·产品特点

1、目前市场最快的操作流程，一致的反应条件完成实验仅需要1小时55分钟； 

2、彻底转换未甲基化的胞嘧啶为尿嘧啶，被修饰的DNA> 99.98%； 

3、修饰过程中具备最低的DNA降解，能阻止超过90%的DNA损失； 

4、启动DNA修饰最低要求：仅需要50pg或者20个细胞；

5、非常简单，可信赖的实验结果和稳定的修饰条件.

·产品说明书

The Methylamp™ DNA Modification Kit is a complete set of optimized reagents designed to convert DNA so that 5-methylcytosines can be detected in downstream applications, for the purpose of gene-specific DNA methylation analysis. The kit has the following advantages and features:

• Total protocol time is less than 2 hours.
• 99.9% conversion rate of unmethylated cytosine into uracil/thymine
• Extremely low degradation of DNA in the modification process, preventing over 90% of DNA loss.
• Requires as little as only 50 pg of input DNA, equivalent to about 20 cells.
• Extremely simple, robust, and reliable sodium bisulfite conversion protocol.
• Suitable for bisulfite sequencing (direct and cloning), next-generation sequencing, pyrosequencing, PCR (real-time, end-point, methylation-specific), COBRA (combined bisulfite restriction analysis), and methylation-based microarrays.
• Highly compatible with Illumina-based workflows.

Background Information
DNA methylation occurs by the covalent addition of a methyl group at the 5-carbon of the cytosine ring, resulting in 5-methylcytosine. DNA methylation is essential for the development of many species and is often associated with many key biological processes as well as gene regulation. There are various methods used to assess DNA methylation states. However, only bisulfite conversion of genomic DNA, followed by PCR amplification, cloning, and sequencing of individual PCR amplimers yields reliable information on the methylation states of individual cytosines on individual DNA molecules. By treating DNA with sodium bisulfite, cytosine residues are deaminated to uracil while leaving 5-methylcytosine intact, providing researchers with distinguishable methylated positions in the DNA sequence during the downstream application. 

Principle & Procedure
DNA is chemically denatured to allow the sodium bisulfite reagent to react 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 chemical and thermophilic degradation of DNA during the sodium bisulfite treatment process. The included non-toxic, modified DNA capture buffer enables the DNA to bind tightly to the column filter, permitting DNA cleaning to be carried out on the column to effectively remove residual sodium bisulfite and salts. Modified DNA can then be eluted for use in various methylation analysis applications or otherwise stored at -20°C for up to 2 months.

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