发布: 2013年11月20日第3卷第22期 DOI: 10.21769/BioProtoc.966 浏览次数: 10625
评审: Tie Liu
相关实验方案
黑吉豆中的绿豆黄化花叶印度病毒(MYMIV)感染、sRNA库建立和鉴定microRNA的深度测序
Anirban Kundu [...] Genotypic Technology
2016年10月20日 10374 阅读
从标记的细胞类型中分离细胞核、提取RNA并去除核糖体RNA以用于RNA-Seq 分析
Mauricio A. Reynoso [...] Kaisa Kajala
2018年04月05日 14848 阅读
Abstract
Medicago truncatula serves as a model plant for legume genetics and genomics. We used RNA-Seq to characterize the transcriptome during the early organogenesis of the nodule and during its functioning. We generated approximately 135.5 million high-quality 36-bp reads, which were then aligned with the M. truncatula genome sequence (Mt3.0 version) and with sequences from a custom splice-junction database, for the detection of transcribed regions and splicing sites. Mapping and analysis of the reads conducted to the detection of 37,333 expressed transcription units (TUs), 1,670 had never been described before and were functionally annotated. We identified 7,595 new transcribed regions, mostly corresponding to 5’ and 3’ UTR extensions and new exons associated with 5,264 previously annotated genes. We also assessed the complexity in the nodulation transcriptome by performing a Cufflinks analysis to determine the frequency of the various alternatively spliced forms. Thus, we identified 23,164 different transcripts derived from 6,587 genes. Finally, we carried out a differential expression analysis, which provided a comprehensive view of transcriptional reprogramming during nodulation.
All Illumina sequence data have been deposited in the NCBI short-read archive, and Sanger-sequenced PCR products have been deposited in GenBank (SRA048731). Assembled contigs longer than 200 bp have been deposited at TSA (JR366937-JR375780). Coverage data are available at http://ddlab.sci.univr.it/cgi-bin/gbrowse/medicago/.
Materials and Reagents
Equipment
Software
Procedure
文章信息
版权信息
© 2013 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Readers should cite both the Bio-protocol article and the original research article where this protocol was used:
分类
植物科学 > 植物分子生物学 > RNA
系统生物学 > 转录组学 > RNA测序
您对这篇实验方法有问题吗?
在此处发布您的问题,我们将邀请本文作者来回答。同时,我们会将您的问题发布到Bio-protocol Exchange,以便寻求社区成员的帮助。
提问指南
+ 问题描述
写下详细的问题描述,包括所有有助于他人回答您问题的信息(例如实验过程、条件和相关图像等)。
Share
Bluesky
X
Copy link