m6A RNA甲基化文库构建试剂盒(测序版) 货号：A-P-9016
EpiNext CUT&RUN RNA m6A-Seq Kit
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背景资料：N6-methyladenosine (m6A) is the most common and abundant modification on RNA molecules present in eukaryotes. Recently, DNA m6A is also identified in multicellular eukaryotes including Caenorhabditis elegans and Drosophila melanogaster, and furthermore identified in higher eukaryotes including plants, mouse and human cells. m6A plays crucial roles in regulating DNA replication, DNA damage, RNA splicing, transposition, transcription, and cellular defense. In human cells, the m6A modification is probably catalyzed by a methyltransferase complex METTL3/METTL14 and removed by the α-ketoglutarate (α-KG)- and Fe2+-dependent dioxygenases such as FTO, ALKBH5 and TET-like enzymes. It was shown that METTL3 and α-KG /Fe2+-dependent dioxygenases play important roles in many biological processes, ranging from development and metabolism to fertility. The dynamic and reversible chemical m6A modification on DNA /RNA may also serve as a novel epigenetic marker of profound biological significance. Down-regulation of m6A modification was first characterized in human cancer cells and tissues, relative to their normal controls. m6A is found to be the most regulated DNA modification in cancers. In addition to the regulation in cancer cells, relative to the primary cell/tissues which contain quite low DNA m6A (<0.001%), a hundreds-fold increase of m6A modification was found for in vitro cultured human cells (0.03%-0.22%)。
产品描述：This kit is designed for measuring total m6A demethylase activity/inhibition. In an assay with this kit, the unique m6A substrate is stably coated on the strip wells. Active m6A demethylases bind to and demethylate m6A contained in the substrate. The un-demethylated m6A in the substrate can be recognized with a high affinity m6A antibody and the immuno-signal is enhanced with enhancer solution. The ratio or amount of un-demethylated m6A, which is inversely proportional to enzyme activity, can then be colorimetrically quantified through an ELISA-like reaction。
产品特点：
1·High enrichment: Use RNA cleavage enzyme mix to simultaneously fragment RNA and cleave/remove any RNA sequences at both ends of the target m6A -containing sequences without affecting RNA regions occupied by the antibody. Short RNA fragments are generated only bound with anti- m6A antibody. True target m6A-enriched regions can, therefore, be reliably identified and high-resolution mapping achieved；
2·Low Input: Unbound RNA cleavage and immunocapture are processed in the same single-tube, which enables the maximal protection of the target m6A-containing regions and the minimized sample loss, allowing the input RNA to be as low as 500 ng；
3.Minimal Background: Cleavage of unbound RNA sequences in the two ends of the target m6A-containing sequences enables the minimized MeRIP/sequencing background, allowing data analysis with <10 million reads.
4.Fast, streamlined procedure: The procedure from RNA to library cDNA is less than 6 hours with <1 h of hands-on time；
5.Highly convenient: The kit contains all required components for each step of the CUT&RUN RNA m6A -Seq, which are sufficient for both m6A-containing RNA sequence capture and captured cDNA library preparation, thereby allowing CUT&RUN RNA m6A –Seq to be the most convenient with reliable and consistent results。
数据分析：
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标准曲线 |
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| EpiNext m6A RNA甲基化文库构建试剂盒(测序版) 流程图 |
Size distribution of library fragments. m6Afragments were enriched from 3 ug of total RNA by the anti- m6A antibody (EpiGentek, #A-1801) and used for cDNA library preparation. Peak at 195 bps reflects insert |
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。
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