版块导航: 正在加载中...

登录注册

应《网络安全法》要求，自2017年10月1日起，未进行实名认证将不得使用互联网跟帖服务。为保障您的帐号能够正常使用，请尽快对帐号进行手机号验证，感谢您的理解与支持！

艾德科技（北京）有限公司

http://www.ivdshow.cn

表观遗传学（m6A RNA甲基化、RNA甲基化、DNA甲基化、DNA羟甲基、DNA甲酰基、DNA羧基），分子生物学，免疫生物学（一抗、二抗），细胞生物学，实验仪器学（核酸提起仪、超微量分光光度计）等等产品与技术服务

北京市昌平区中关村生命科学园东60米艾德园

联系人: Peter Chen / 邮箱: 1951545998@qq.com / QQ: 1513545070 / 电话: 0313-5935521 / 手机: 18911529660

关注收听

返回列表

【有奖交流】积极回复本帖子，参与交流，就有机会分得作者 ADTechnology 的 75 个金币，回帖就立即获得 1 个金币，每人有 1 次机会

ADTechnology

捐助贵宾 (著名写手)

表观遗传学专业供应商

应助: 12 (小学生)
金币: 12559.7
散金: 1726
红花: 1
帖子: 2511
在线: 44.6小时
虫号: 1787705
注册: 2012-04-30
专业: 人类遗传学

[商家供应] 通过qPCR来检测5mC和5hmC含量---双位点5mC&5hmC定量检测试剂盒--(A-P-1067)

本商家没有参加优惠活动，一起邀请他来参加吧我的代金券

代金券0元

我要兑换代金券

限量 x 张
满 x 可使用；有效期 x 天；过期可退

内容

双位点5mC&5hmC定量检测试剂盒又名:特定位点甲基化和羟甲基化qPCR定量检测试剂盒货号：A-P-1067

BisulPlus Loci 5mC & 5hmC Detection PCR Kit

背景资料：DNA methylation occurs by the covalent addition of a methyl group at the 5-carbon of the cytosine ring, resulting in 5-methylcytosine (5mC). In somatic cells, 5mC is found almost exclusively in the context of paired symmetrical methylation of the dinucleotide CpG, whereas in embryonic stem (ES) cells, a substantial amount of 5mC is also observed in non-CpG contexts. The biological importance of 5mC as a major epigenetic modification in phenotype and gene expression has been widely recognized. 5-hydroxymethylcytosine (5hmC), as a sixth DNA base with functions in transcription regulation, has been detected to be abundant in human and mouse brains and embryonic stem (ES) cells. In mammals, it can be generated by the oxidation of 5mC, a reaction mediated by the ten-eleven translocation (TET) family of 5mC-hydroxylases. 5hmC has been demonstrated to be tissue specific, ranging from undetectable levels in cultured cell lines to 0.6% in human brain tissues, and can be as high as 8% of total DNA in some other species. The biological significance of 5hmC as an important epigenetic modification in phenotype and gene expression has been recently recognized. For example, global decrease in 5hmC content (DNA hypohydroxymethylation) has been exhibited in nearly all cancers and has been proposed as a molecular marker and therapeutic target in cancer as well. More importantly, in contrast to 5mC that is generally considered as a gene repression marker, 5hmC is regarded as an intermediate of DNA demethylation and associated with expressed genes with a potential role in activating genes. Like 5mC, 5hmC remains the same after bisulfite-conversion of cytosine to uracil (C to U), which makes 5hmC indistinguishable from 5mC. As consequence, all 5hmC will be misread as 5mC in the assay with using MS-PCR or bisulfite-seq, which results in that the identified DNA methylation status is incorrect as it in fact includes both 5mC and 5hmC。

产品描述：This kit includes all reagents required for successful qPCR using converted DNA generated from a tiny amount of input DNA. In this preparation, DNA is simultaneously bisulfite modified and fragmented to the appropriate length during the bisulfite process. During the bisulfite treatment, unmodified cytosine (C) is converted to uracil and will be read as T in the sequencing. 5-methylcytosine (5mC) remains the same and 5-hydroxymethylcytosine (5hmC) forms cytosine 5-methylenesulfonate (CMS). The bisulfite modified DNA is further treated with specific APOBEC deaminase, which converts 5mC to thymine but not affect CMS. During the PCR and sequencing CMS will still be read as C so that the 5hmC can be discriminated not only from C, 5mC but also other modified cytosines such as 5fC and 5caC (see the table 1). The bisulfite-enzyme converted DNA can then be used for qPCR for loci specific detection of 5mC and 5hmC。

产品特点：

1·Simultaneous Reading

NA treatment is prepared simultaneously for both 5hmC and 5mC, which allows for parallel identification in a loci- or gene-specific manner simultaneously by qPCR；

2·Innovative Method:A combination of bisulfite and subsequent cytosine deaminase (APOBEC) treatment allows for tactical conversion of 5mC in order to identify 5hmC；

3.High Specificity:5hmC is discriminated from C and 5mC as well as from other modified cytosines 5fC and 5caC.

4.Flexibility: Perform 24 reactions for 5hmC and 24 reactions for 5mC analysis simultaneously, or perform 48 reactions for just 5mC；

5.Fast and Streamlined Procedure:The procedure from DNA bisulfite treatment to PCR products can be completed within 5 hours；

6.Complete Conversion: Completely converts C into uracil (>99%) and 5mC to thymine (>95%)；

7.Broad sample suitability:Starting materials can be genomic DNA isolated from various tissue/cell samples such as fresh and frozen tissue, cultured cells from a flask or microplate, microdissection samples, paraffin-embedded tissue, biopsy, embryonic cells, plasma/serum samples, and body fluid samples, etc. DNA enriched from various enrichment reactions such as ChIP, MeDIP/hMeDIP, or exon capture may also be used as starting material.

8.Simple, reliable, and consistent assay conditions。

数据分析：

操作流程

Discrimination of 5hmC from cytosine and 5mC by qPCR: Unmodified, methylated and hydroxymethylated DNA standards are used for bisulfite-enzyme treatment using the he BisulPlus™ Loci 5mc & 5hmC Detection PCR Kit followed by real-time amplification with use of primers targeting to CpG converted or unconverted sequences: A: BisulPlus; B: Bisulfite standalone. Amplification lines are highlighted for visibility.

参考文献：

1.Lewinska A et. al. (February 2017). Downregulation of Methyltransferase Dnmt2 Results in Condition-dependent Telomere Shortening and Senescence or Apoptosis in Mouse Fibroblasts. J Cell Physiol.

保存建议：在接收到艾德寄出的试剂盒后请按照说明书建议，使用不同的保存条件或温度来保存试剂盒内组分。
更多请参考官网:猛戳产品详情