在本项研究中，作者仅使用了2.5 μg 的骨髓瘤临床样品total RNA（direct RNA-seq 推荐量：50 μg）用于xPore m6A位点检测，利用METTL3-KO HEK293T细胞作为未修饰对照进行xPore模型建立，克服了临床样品缺乏相应对照组的问题。综上，该研究开发了一种基于Nanopore direct RNA测序的RNA甲基化差异分析计算方法，实现...
1. 什么是Nanopore 测序 测序大家都并不陌生,通过RNA-seq等测序技术我们了解了很多调控的信息。但是这些测序往往是短读长(short read)或者是cDNA测序的结果,对于一些链特异性的结果,我们需要通过对应的测序技术等等。nanopore是第三代的测序技术,主要通过在nanopore周围添加电解液,电压驱动以及驱动蛋白牵引带电粒子的方式...
Nanopore is the only technology that can directly sequence RNA strands without reverse transcription or PCR reactions. Nanopore direct RNA sequencing technology sequences natural RNA, allowing it to retain and detect RNA base modification information, estimate the poly(A) tail length relatively accurately...
参考文献: [1]Parker M T , Knop K , Sherwood A V , et al. Nanopore direct RNA sequencing maps the complexity of Arabidopsis mRNA processing and m6A modification[J]. Elife Sciences, 2020, 9:e49658. [2]Froussios K , Kira Mourão, Simpson G , et al. Relative Abundance of Trans ( ...
本周一,Nature Methods在线发表Oxford Nanopore direct RNA测序技术测评文章,结果表明,【不经反转、无须扩增的RNA直接测序】能获得全长的链特异性RNA,无测序偏好性,并同时记录碱基修饰,为后续研究基因结构和基因表达,提供新技术新方法。 direct RNA建库测序
1. 作为长读长测序技术,能减少短读长测序打断后重新拼接带来的歧义,结果准确度更高、更可信; 2. 能够直接对RNA进行测序,避免逆转录和PCR扩增导致的RNA修饰信息缺失; 3. 可以直接检测m6A等碱基修饰,配合优化的算法,获得无损的m6A单碱基分辨率图谱; 4. 一次测序,多套数据:转录组图谱、m6A修饰图谱、异构体、可变剪...
该研究应用Nanopore Direct RNA测序技术研究中华鳖转录组的复杂性。与M表型相比,F表型可变剪接差异显著。m5C和m6A两种RNA甲基化修饰在不同性别表型之间存在差异,并对关键基因的m6A甲基化水平进行了验证。该研究揭示了生物表观遗传在中华鳖性逆转过程中扮演的重要作用。
近日,福建农林大学海峡联合研究院林学中心顾连峰教授研究组在Plant Physiology发表了题为“Drought induces epitranscriptome and proteome changes in stem-differentiating xylem of Populus trichocarpa”的研究论文,揭示了毛果杨次生木质部中RNA全长转录本比率、N6-甲基腺嘌呤(N6-methyladenosine, m6A)修饰率、Poly(A)位点...
Standard direct RNA-sequencing (RNA-seq) relies on poly(A) selection by using a poly(T)-tethering adaptor that base-pairs canonically to a motor protein linked adaptor, facilitating the movement of the polynucleotide string through the protein pore (Fig.1A). While standard direct RNA-seq provid...
Characterizing complex viral transcriptomes by conventional RNA sequencing approaches is complicated by high gene density, overlapping reading frames, and complex splicing patterns. Direct RNA sequencing (direct RNA-seq) using nanopore arrays offers an e