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Epigenetics |
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| Epigenetics>【Zymo】 |
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Quest 5-hmC™ DNA Enrichment Kit(D5420) |
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Quest 5-hmC™ DNA Enrichment Kit
About Quest 5-hmC™ DNA Enrichment Kit
While the importance of DNA methylation in epigenetic regulation is well established, the biological role of hydroxymethylation remains elusive. The “sixth base”, 5-hydroxymethylcytosine (5-hmC), has been detected in the DNA of embryonic stem cells and other cell types. Brain tissue DNA contains the highest levels of 5-hmC. Recent work suggests that 5-hmC may function in gene regulation and may be involved as an intermediate in active demethylation of 5-methylcytosine (5-mC). The Quest 5-hmC™ DNA Enrichment Kit features J-base binding protein (JBP) for the specific enrichment of 5-hmC containing DNA. The consolidated workflow makes the procedure reliable for robust analysis of multiple samples. Simply glucosylate the input DNA, add JBP Capture MagBeads, and then wash and elute the enriched 5-hmC DNA.
| Equipment |
Microcentrifuge, heat block, incubator, and a rotator/rocker are required. A magnetic stand is recommended, but this kit includes magnetic rods if such a stand is unavailable. A thermal cycler will be necessary to perform PCR. |
| Sample Sources |
This protocol is developed for processing fragmented genomic DNA as the input material. DNA should be free of enzymatic inhibitors and can be in water, TE, or low salt buffer. |
| Input |
5-4,000 ng DNA |
| Purity |
Enriched 5-hmC DNA is of suitable purity for a wide variety of downstream applications including PCR, qPCR, Next-Gen sequencing, whole genome amplification, arrays, and more. |
| 5-hmC Glucosyltransferase |
One unit (1 U) of recombinant 5-hmC Glucosyltransferase (5-hmC GT) is sufficient to glucosylate 1 μg of 5-hmC containing DNA in 1 hour at 37 °C. |
Clean & uniform enrichment of 5-hmC DNA by J-Binding Protein (JBP-1).
Simple three-step workflow
Enriched DNA is ideal for PCR, qPCR, Next-Gen sequencing, arrays, and more.
| Quest 5-hmC™ DNA Enrichment Kit |
25 Rxns. |
D5420 |
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| Quest 5-hmC™ DNA Enrichment Kit |
50 Rxns. |
D5421 |
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表观遗传学产品全面解决方案列下：
推荐阅读：
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-羟甲基化胞嘧啶，并且区分5-hmC, 5-mC, 和 C，使得研究者能够重新评估他们的DNA甲基化信息，也能够在新样品中寻找DNA羟甲基化。 |
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