高敏感亚硫酸盐测序试剂盒(illumina) 货号:A-P-1056
EpiNext Bisulfite-Seq High Sensitivity Kit (Illumina)
背景资料:carbon of the cytosine ring, resulting in 5-methylcytosine (5-mC). DNA methylation is essential in regulating gene expression in nearly all biological processes including development, growth, and differentiation. Alterations in DNA methylation have been demonstrated to cause a change in gene expression. Genome-wide analysis of DNA methylation could provide valuable information for discovering epigenetic markers used for disease diagnosis, and potential targets used for therapeutics. Bisulfite sequencing via next-generation sequencing technologies allow for high volume, lower cost output of DNA sequence data towards a better understanding of DNA methylation.
产品描述:This kit includes all reagents required for a successful preparation of a DNA library by directly using bisulfite-converted DNA generated from a small amount of input DNA (500 pg to 500 ng). In this preparation, DNA is bisulfite converted and fragmented to the appropriate length simultaneously during the bisulfite process. The bisulfite-treated DNA, which is in single stranded form, is then converted to dsDNA and directly used for ligation with BisDNA-specific adaptors that are necessary for amplification and sequencing. The fragments are size selected and purified using MQ binding beads, which allows for quick and precise size selection of DNA. Size-selected DNA fragments are amplified using a high-fidelity PCR Mix which ensures maximum yields from minimum amounts of starting material and provides highly accurate amplification of library DNA with low error rates and minimal bias。
产品特点:
1·Innovative method - Allows for simultaneous bisulfite conversion and size-appropriate DNA fragmentation. The bisulfite DNA can be directly ligated to adaptors thereby eliminating the possibility of breaking adaptor-ligated fragments, which often occurs with traditional WGBS and RRBS methods;
2·Fast and streamlined procedure - the procedure from DNA bisulfte treatment to PCR amplification can be finished within the same day (<8 h). Gel-free size selection/purification saves time and prevents handling errors, as well as loss of valuable samples;
3·Complete conversion - The innovation reagent composition converts unmethylated cytosine into uracil at a level greater than 99.9%, with negligible inappropriate- or error-conversions of methylcytosine to thymine (<0.1%);
4·High sensitivity and efficiency - Adaptor ligation after bisulfite treatment eliminates loss of fragments and selection bias, which enables input DNA to be as low as 0.5 ng;
5·Extremely convenient - the kit contains all the required components for each step of the DNA library preparation process, which are sufficient for bisulfite conversion, ligation, clean-up, size selection, and library amplification, thereby allowing the bisulfite DNA library preparation to be streamlined for the most reliable and consistent results;
6.Minimal bias - Ultra HiFi amplification enables achievement of reproducibly high yields of DNA libraries with minimal sequence bias and low error rates;
7.Broad sample suitability - Starting materials can be genomic DNA isolated from various tissue/cell samples such as fresh and frozen tissues, cultured cells from a flask or microplate, microdissection samples, paraffin-embedded tissues, biopsy samples, 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 materials。
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
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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。 |