K1501 Thermo EpiJET 5-hmC和 5-mC 分析试剂盒,EpiJET 5-hmC and 5-mC Analysis Kit /EpiJET 5-hmC和 5-mC 分析试剂盒-试剂- bio-rad伯乐一级代理 |伯乐Aminex |伯乐电泳槽1658001| 伯乐ddPCR bio-rad授权代理商、biorad伯乐qpcr仪

K1501 Thermo EpiJET 5-hmC和 5-mC 分析试剂盒,EpiJET 5-hmC and 5-mC Analysis Kit /EpiJET 5-hmC和 5-mC 分析试剂盒

2024-10-24

EpiJET 5-hmC and 5-mC Analysis Kit /EpiJET 5-hmC和 5-mC 分析试剂盒

货号：K1501

规格：25 reactions

价格：2313

产品类型：表观试剂

品牌：Thermo Fisher

Themo Scientific EpiJET 5-hmC and 5-mC Analysis Kit includes three advanced enzymes: T4 phage β-glucosyltransferase (T4 BGT), Epi MspI and Epi HpaII. The Epi MspI and Epi HpaII are isoschisomers with differing sensitivity to internal C methylation of the CCGG site. Epi MspI completely digests genomic DNA when cytosine, 5-methylcytosine (5-mC) or 5-hydroxymethylcytosine (5-hmC) is within its recognition sequence, but its activity is completely blocked when 5-hmC is glucosylated. Epi HpaII cleaves the unmodified CCGG site only.The kit takes advantage of the T4 BGT ability to specifically modify 5-hmC residues by adding glucose moiety to 5-hmC and the sensitivity of restriction enzyme Epi MspI to glucosylated 5-hmC. The T4 BGT is formulated for highly specific, complete and fast glucosylation of 5-hmC in 15 minutes.The analysis involves DNA sample glucosylation and subsequent restriction digestion with Epi MspI and Epi HpaII. Restriction digestion products are then analyzed by qPCR with a primer pair flanking CCGG site of interest.

优点：

-Efficient and reliable—accurate quantification of 5-hmC and 5-mC-Robust—broad range (from 0.2 µg to 4 µg) of input DNA-Fast—procedure completed in 5 hours, no overnight incubation.

数据：

图1.No cleavage 无切割If Cq1≈ Cq2 ≈ Cq3 ≈ Cq4*,No cleavage.No Epi MspI/HpaII recognition site or CCGG sequence is modified at the external C (Epi MspI/HpaII cleavage is blocked).

图2.No cytosine modification 无胞嘧啶修饰If Cq2 ≈ Cq3 ≈ Cq4 and (Cq2 ≈ Cq3 ≈ Cq4) – Cq1 ≥ 4.5,No cytosine modification.CCGG site does not contain 5-hmC or 5-mC bases. Epi HpaII and Epi MspI are able to cleave CCGG.

图3.Partial cytosine modification 部分胞嘧啶修饰If Cq1 < Cq2 < Cq3 < Cq4,Partial cytosine modification.Cleavage levels represent modification level in the CCGG site of interest. Calculate into 5-hmC, 5-mC and C percentage.

图4.Full hydroxymethylation全部羟甲基化If Cq1 ≈ Cq2 ≈ Cq3 andCq4 - (Cq1 ≈ Cq2 ≈ Cq3) ≥ 4.5,Full hydroxymethylation.Glucosylation blocked cleavage by Epi MspI. CCGG site is fully 5-hydroxymethylated.

图5.Full methylation 全部甲基化If Cq3 ≈ Cq4, Cq1 ≈ Cq2 and (Cq3 ≈ Cq4) – (Cq1 ≈ Cq2) ≥ 4.5,Full methylation.Methylation blocked the cleavage by Epi HpaII. CCGG site is fully 5-methylated.

组成成分：

▪ T4 ß – glucosyltransferase, 5 U/µL▪ 10X Epi Buffer▪ 10X UDP–glucose▪ Epi MspI▪ Epi HpaII▪ 5-hmC Control DNA▪ 5-mC Control DNA▪ Unmodified Control DNA▪Control primer 1▪ Control primer 2▪ Water, nuclease-free▪ Detailed Protocol

K1501 Thermo EpiJET 5-hmC和 5-mC 分析试剂盒,EpiJET 5-hmC and 5-mC Analysis Kit /EpiJET 5-hmC和 5-mC 分析试剂盒

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