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填写我的在线表格。如何选择 DNA 和 RNA 纯化试剂盒选择满足您需求的 DNA 或 RNA 纯化试剂盒阅读更多 A First Look at A NEANDERTHAL FACE DNA 甲基化分析和现代人类的进化史阅读更多如何获得用于芯片测序的高质量 DNA 成功 ChIP 的必备技巧-Seq阅读更多如何在 ILLUMINA® 平台上对 RRBS 文库进行测序了解如何在 Illumina® 平台上对 RRBS 文库进行测序阅读更多从生物化学到酿造的酵母

$\"来自生物化学到酿造\"$ 由生物化学家转型为酿酒师的酵母问答阅读更多主题COVID-19DNA环境表观遗传学微生物组NGSP质粒纯化RNA样品集合酵母您的研究有一个故事。让我们分享。

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DNA Hydroxymethylation

$\"\"$

PRODUCT OVERVIEW

QUEST 5-hmC DETECTION KIT™

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 an \"ultra” modified base, glucosyl-5-hydroxymethylcytosine (Figure 1).

$\"Hydroxymethylcytosine$

Figure 1: 5-hmC Glucosyltransferase transfers a glucose moiety from uridine diphosphoglucose (UDPG) onto preexisting 5-hydroxymethylcytosines within DNA.

After glucosylation of 5-hydroxymethylcytosine, digestion of DNA with GSRE (glucosyl-5-hydroxymethylcytosine sensitive restriction endonucleases, such as GlaI, MspI and TaqI), allows for effective differentiation of 5-methylcytosine from 5-hydroxymethylcytosine. Identification of 5-hydroxymethylcytosine in a sequence specific context can then be deduced from the restriction endonuclease recognition sequence.

$\"Quest$
Click image for larger view

Figure 2: Quest 5-hmC Detection Kit™ Workflow Treatment of DNA containing 5-hmC with 5-hmC Glucosyltransferase specifically adds a glucose moiety, yielding glucosyl-5-hydroxymethylcytosine. Subsequent GSRE (glucosyl-5-hydroxymethylcytosine sensitive restriction endonucleases, such as MspI and GlaI) digestion, will cut DNA with 5-methylcytosine or 5-hydroxymethylcytosine in their recognition sequence, but will be blocked by the glucosyl-5-hydroxymethylcytosine DNA.

SCIENTIFIC FOCUS

5-HYDROXYMETHYLCYTOSINE

What is 5-hydroxymethylcytosine?
5-hydroxymethylcytosine (5-hmC) is the hydroxylated form of the well-known 5-methylcytosine (5-mC). It was first discovered in bacteriophage DNA in 1952[1]. During the next several decades, work on hydroxymethcytosine focused primarily on its effects on protecting phage DNA. It is only more recently that attention has turned to its role in the development of human and mouse brain, as well as embryonic stem cell[2], where hydroxymethylcytosine was found to widely exist and can be generated by oxidation of the 5-mythylcytosine catalyzed by a enzyme called Tet[3].

The 5-hmC is also referred to as the 6th base of DNA in reference to the 5-methylcytosine as the 5th base by some authors. The 5-hmC can be further modified by certain glucose transferase at the hydroxyl position to form glucosyl-5-hydroxymethylcytosine. See the structure of cytosine and its naturally occuring derivatives below.

$\"What$

The exact function of this modified base is still under investigation, but it is thought to be involved in DNA demethylation or gene expression regulation. However, the lack of a valid technique which can accurately detect 5-hmC sensitively has hindered researchers to further explore its potential biological function. This results from the fact that 5-hmC behaves very similarly to its precursor 5-mC in bisulfite conversion and restriction enzyme digestion[4, 5]. Zymo Research overcame this obstacle and is introducing the first kit available for sequence specific determination of 5-hmC, Quest 5-hmC Detection Kit™.

References
[1] Wyatt GR, Cohen SS(1952) . A new pyrimidine base from bacteriophage nucleic acids. Nature 170 (4338) 1072-3.
[2] Kriaucionis S, Heintz N (2009). The nuclear DNA base 5-hydroxymethylcytosine is present in Purkinje neurons and the brain. Science 324 (5929): 929-30.
[3] Tahiliani M et al. (2009). \"Conversion of 5-methylcytosine to 5-hydroxymethylcytosine in mammalian DNA by MLL partner TET1\". Science 324 (5929): 930–35
[4] Huang Y.,et al (2010) The Behavior of 5-Hydroxymethylcytosine in Bisulfite Sequencing PLoS One.; 5(1) e8888
[5]Jin S et al., (2010) Examination of the specificity of DNA methylation profiling techniques towards 5-methylcytosine and 5-hydroxymethylcytosine Nucleic Acids Res. 38(11): e125.

Quest 5-hmC Detection Kit™ and Quest 5-hmC Detection Kit™ - Lite contain patent pending technologies.

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