EpiNext RNA甲基化极易测序分析试剂盒（illumina仪器配套专用） 货号：A-P-9007
EpiNext 5-mC RNA Bisulfite-Seq Easy Kit (Illumina)
背景资料：5-methylcytosine (5-mC) in DNA occurs by the covalent addition of a methyl group at the 5-carbon of the cytosine ring by DNA methyltransferases. This process has been well studied and is generally associated with repression of gene expression. It was also observed that in humans, 5-mC occurs in various RNA molecules including tRNAs, rRNAs, mRNAs and non-coding RNAs (ncRNAs). At least 10,275 5-mC candidate sites were discovered in mRNAs and ncRNAs, which cover 10.6% of the total cytosine residues in the transcriptome. 5-mC seems to be enriched in some classes of ncRNA, but relatively depleted in mRNAs. However, the majority (83%) of their candidate sites were found in mRNAs. Within these transcripts 5-mC appears to be depleted within protein coding sequences, but enriched in 5’ and 3’ UTRs. Two different methyltransferases, NSUN2 and Dnmt2 are known to catalyze 5-mC modification in eukaryotic RNA. Recent data strongly suggest that RNA cytosine methylation affects the regulation of various biological processes such as RNA stability and mRNA translation. Furthermore, loss of 5-mC in vault RNAs causes aberrant processing into Argonaute-associated small RNA fragments that can function as microRNAs. Thus, impaired processing of vault ncRNA may contribute to the etiology of human disorders related to NSun2-defciency 。
产品描述：This kit includes all reagents required for a successful RNA bisulfite conversion and bisulfite RNA library preparation using bisulfite-converted RNA generated from a wide range of input RNA amounts (5 ng to 500 ng). In this preparation, RNA is simultaneously bisulfite converted and fragmented to the appropriate length during the bisulfite process. The bisulfite-treated RNA, which is in single stranded form, is then simultaneously converted to double stranded cDNA and adaptor tagged. The tagged fragments are size selected and purified using MQ binding beads, followed by amplification with a high-fidelity PCR Mix, which ensures maximum yields from minimum amounts of starting material and provides highly accurate amplification of library cDNA with low error rates and minimum bias。
产品特点：
1·High sensitivity and efficiency: By using an innovative method that enables adaptor tagging of bisulfite-converted RNA to be intact through random probing with blocking formation of un-tagged or half-tagged RNA fragments, both non-barcoded (singleplexed) and barcoded (multiplexed) library preparation can be reliably made with high sensitivity and efficiency, thereby providing a robust and reliable means to build 5mC RNA-seq library. The optimized RNA bisulfite method and enhanced adaptor tagging eliminates loss of fragments and selection bias, which enables input RNA to be as low as 5 ng with increased efficiency of completely tagged cDNA library；
2·Easy and streamlined procedure: The entire procedure can be finished in 6 hours. Gel-free size selection/purification saves time and prevents handling errors, as well as loss of valuable samples；
3.Complete conversion: The innovative reagent composition converts unmethylated cytosine into uracil at a level greater than 99.9%, with no or negligible inappropriate/error conversion of methylcytosine to thymine (<0.1%) when the indicated range of sample RNA is used；
4.Extremely convenient: The kit contains all the required components for each step of the RNA library preparation process, which is sufficient for bisulfite conversion, ligation, clean-up, size selection, and library amplification, thereby allowing the bisulfite RNA library preparation to be streamlined for the most reliable and consistent results；
5.Minimal bias: Ultra HiFi amplification enables achievement of reproducibly high yields of bisulfite converted RNA libraries with minimal sequence bias and low error rates；
6.Broad sample suitability: Starting materials can be total RNA isolated from various tissue/cell samples such as fresh and frozen tissue, cultured cells from a flask or microplate。
数据分析：
| 操作流程 |
标准曲线 |
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| EpiNext RNA甲基化极易测序分析试剂盒（illumina仪器配套专用）步骤 |
文库片段大小尺寸：使用EpiNext RNA甲基化极易测序分析试剂盒（illumina仪器配套专用）对于输入的50ng小鼠RNA Post-bisfulte后的cDNA文库。 |
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。
定购信息：
| 产品名称 |
规格 |
操作手册 |
货号 |
询价 |
| EpiNext RNA甲基化极易测序分析试剂盒（illumina仪器专用） |
24次 |
英文PDF-中文PDF |
A-P-9007-24 |
 |
| EpiNext RNA甲基化极易测序分析试剂盒（illumina仪器专用） |
12次 |
英文PDF-中文PDF |
A-P-9007-12 |
|
| 总体RNA甲基化极易定量检测试剂盒(荧光法) |
96次 |
英文PDF-中文PDF |
A-P-9009-96 |
| 总体RNA甲基化极易定量检测试剂盒(荧光法) |
48次 |
英文PDF-中文PDF |
A-P-9009-48 |
|
| 总组蛋白H3K27乙酰化定量检测试剂盒（比色法） |
48次 |
PDF |
A-P-4059-48 |
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| 总组蛋白H3K27乙酰化定量检测试剂盒（比色法） |
96次 |
PDF |
A-P-4059-96 |
|
| EpiNext 高灵敏重亚硫酸盐测序试剂盒（Illumina） |
12次 |
PDF |
A-P-1056A-12 |
| EpiNext 高灵敏重亚硫酸盐测序试剂盒（Illumina） |
24次 |
PDF |
A-P-1056A-24 |
|
| 96孔DNA甲基化极速修饰试剂盒（磁珠法） |
96次 |
英文PDF-中文PDF |
A-P-1050-096 |
|
| 96孔DNA甲基化极速修饰试剂盒（磁珠法） |
192次 |
英文PDF-中文PDF |
A-P-1050-192 |
|
| 抗人C5a/C5a desArg单克隆抗体(克隆号2942) |
100ug |
|
C-HM2078 |
|
| 抗人TNF-R II单克隆抗体(克隆号80M2),FITC标记 |
100ug |
|
C-HM2022F |
|
| 抗人TNF-R II单克隆抗体(克隆号MR2-1),生物素标记 |
50ug |
|
C-HM2008 |
|
| 抗人TNF-R I单克隆抗体(克隆号MR1-2),生物素标记 |
50ug |
|
C-HM2006 |
|
| 抗小鼠TREM-2单克隆抗体(克隆号6E9) |
100ug |
PDF |
C-HM1129F |
|
| m6A RNA甲基化定量检测试剂盒(比色法) |
96 次 |
英文PDF--中文PDF |
A-P-9005-96 |
|
| RNA甲基化修饰试剂盒 |
50 次 |
英文PDF--中文PDF |
A-P-9003-050 |
|
| EZ RNA甲基化修饰试剂盒 |
50 次 |
英文PDF--中文PDF |
A-R5001 |
|
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。 |
