Quest 5-hmC Detection Kit™ - Lite 货号：A-D5415
背景资料：The Quest 5-hmC Detection Kit™ from Zymo Research allows for sequence specific detection of 5-hydroxymethylcytosine within DNA using a simple and efficient reaction setup. Utilizing a robust and highly specific 5-hmC Glucosyltransferase enzyme, 5-hydroxymethylcytosine in DNA is specifically tagged with a glucose moiety yielding a modified base, glucosyl-5-hydroxymethylcytosine (Figure 1).
After glucosylation of 5-hydroxymethylcytosine, digestion of DNA with glucosyl-5-hydroxymethylcytosine sensitive restriction endonucleases (GSREs) allow differentiation of 5-methylcytosine from 5-hydroxymethylcytosine according to the context of a GSRE's recognition sequence (see Table 1). GSRE's can efficiently digest DNA when cytosine, 5-methylcytosine, or 5-hydroxymethylcytosine is within their recognition sequence. However, if 5-hydroxymethylcytosine is glucosylated (i.e., glucosyl-5-hydroxymethylcytosine), GSREs can no longer digest the DNA. Therefore, by exploiting this sensitivity to glucosyl-5-hydroxymethylcytosine, effective detection of 5-hydroxymethylcytosine can be achieved by a number of downstream applications (e.g.- qPCR, NextGen Sequencing, Southern blotting, microarray, etc.). QPCR is recommended to assess locus-specific context of 5-hydroxymethylcytosine within the DNA (see Figure 3). Controls and protocols for qPCR 5-hydroxymethylcytosine detection are included with the kit.
产品描述：Method to distinguish 5-hydroxymethylcytosine in sequence- and locus-specific context within DNA.
Convenient and reliable single tube reaction format.
DNA is eluted in water or low salt buffer and is suitable for analysis by a variety of downstream applications.
产品特点：
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5-hmC Glucosyltransferase
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5-hmC Glucosyltransferase is guaranteed for 1 year at -20°C. Long term storage at -80°C is recommended. Avoid multiple freeze thawing. |
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Quest 5-hmC Control DNA
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A 90 bp DNA standard used for qPCR. [10 ng/μl] in storage buffer (10mM Tris-HCl; 0.1mM EDTA; pH 8.0) supplied in 30 μl volume. See Appendix I (pg. 6) in Protocol for more details. |
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Quest qPCR Primers
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Control primers specific to Quest 5-hmC Control DNA validated for qPCR. See Appendix II (pgs. 7, 8) in Protocol for detailed information. |
保存建议：请在接收到Quest 5-hmC Detection Kit™ - Lite 产品后，按照产品操作说明书对试剂盒内不同组份进行适当的保存。
定购信息：
| 产品名称 |
规格 |
操作手册 |
货号 |
询价 |
| Quest 5-hmC Detection Kit™ - Lite |
25 次 |
PDF |
A-D5415 |
 |
| Quest 5-hmC Detection Kit™ - Lite |
50 次 |
PDF |
A-D5416 |
|
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关于5-hmC
5-hmC是近年来在动物组织中发现的，由胞嘧啶修饰而来。5-hmC在表观遗传学上的功能可能与5-甲基化胞嘧啶(5-mC)不同。尽管到现在为止还不确知其功能，有研究者猜测它在调控基因的表达与关闭过程中起着重要的作用。5-mC的发现让我们不得不重新评估DNA甲基信息，也不得不监测人类的健康组织和病理组织之间5-mC相对分布的差异。在EPI公司的MethylFlash技术之前，我们还没有发现任何直接的常规方法来检测5- hmC,以及区分5-hmC和5-mC
5-hmC 和 5-mC的区别
时下常用的DNA甲基化分析方法包括限制内切酶酶切和亚硫酸氢盐或MeDIP介导的MS-PCR和测序，这些技术都不适合用来检测5-hmC，因为它与5-mC事实上很难用这类方法区分开来。为了解决这个问题，EPIk研制了MethylFlash羟甲基化DNA定量试剂盒(荧光法)。本试剂盒提供了一种很经济的方法来检测5-羟甲基化胞 |
