甲基化 和 RNA甲基化 甲基转移酶促进的过程，其中甲基（CH3）被添加到DNA或RNA分子上的特定原子上。在细胞RNA中，已经鉴定出100多种化学修饰。各种RNA甲基化修饰包括m6A RNA甲基化、m5C RNA甲基化，m1A RNA甲基化和m7G RNA甲基化等。其中，最突出和最广泛研究的是m6A RNA甲基化，发生在RNA分子中腺嘌呤残基的第六个氮原子处（N6甲基腺苷，m6A）。这种修饰代表了真核mRNA中最常见的转录后修饰，约占所有RNA甲基化修饰的80%。
MeRip-seq基于抗体特异性结合甲基化碱基的原理。它利用m6A特异性抗体进行免疫沉淀，以富集经过甲基化的RNA片段。然后采用高通量测序来检测m6A修饰的转录本。然而，这种方法只能识别高甲基化的区域，在识别RNA甲基化方面缺乏单碱基分辨率。
Nanopore sequencing is a long read sequencing technology based on electric signal recognition of base sequences. Different modifications on RNA bases cause variations in the obstruction size as they pass through a nanopore channel, generating characteristic electrical signals. Real-time monitoring of these signals allows for the determination of the corresponding base types and whether they carry base modifications. In other words, Nanopore sequencing can directly perform full-length transcriptome sequencing on natural RNA samples without the need for specific antibody binding. It enables the detection of m6A (N6-methyladenosine) methylation modifications on RNA with single-base resolution, while simultaneously providing quantitative gene/transcript expression and transcript structure identification.
纳米孔测序技术利用检测单个分子通过纳米孔时产生的信号，在孔的两侧产生电位差。纳米孔的直径只允许单个核苷酸聚合物通过。不同碱基（包括具有甲基化修饰的碱基）的带电性质各不相同，从而能够通过电信号的差异检测相应的碱基类型和甲基化信息。
Single Nucleotide Resolution Across the Entire Transcriptome: Identification of m6A methylation modifications at single nucleotide resolution throughout the transcriptome.
Direct Reading of m6A Methylation Information: Direct extraction of m6A methylation information without the need for immunoprecipitation enrichment experiments.
Simultaneous Quantification of Gene/Transcript Expression and Transcript Structure: Conduction of both quantitative analysis of gene/transcript expression and investigation of transcript structure concurrently.
Absence of Reverse Transcription and PCR Bias: Nanopore direct RNA sequencing eliminates the need for disruption, reverse transcription, and PCR amplification, allowing for the direct full-length transcriptome sequencing of natural RNA samples.
(Nicky Jonkhout et al,. 核糖核酸 2017)
m6A RNA甲基化检测：
不同基序间m6A甲基化水平的比较分析。
不同样本间m6A甲基化水平的比较分析。
m6A甲基化水平的功能研究。
基因/转录表达的量化和转录结构变异的鉴定：
在转录水平上准确定量基因/转录物表达，以鉴定差异表达的转录物。
功能基因的鉴定。
精确检测可变剪接、选择性聚腺苷酸化（APA）、融合基因等。
相关性分析：
m6A RNA甲基化与基因/转录物表达的相关性分析。
