Long-read nascent RNA sequencing from S. pombe wild-type and prp2-1 strains

Long-read SMRT cDNA sequencing of nascent RNA from exponentially growing S. pombe 972h- and prp2-1 cells first at 26C and then for 2 hours at 37C was employed to obtain splicing information from single transcripts. Nascent RNA was prepared from the yeast chromatin fraction (Carrillo Oesterreich, Preibisch, Neugebauer, Mol Cell 2010). The nascent 3’ end was labeled with a 3’ DNA adaptor through ligation. The adaptor sequence served as template for full-length reverse transcription and double-stranded cDNA was obtained in a transcriptome-wide PCR. The dataset contains data from 3 replicates per strain. Scripts for demultiplexing the samples can be found on my Github account (https://github.com/lherzel).

本研究采用长读长单分子实时（SMRT）cDNA测序技术，对指数生长期的粟酒裂殖酵母（Schizosaccharomyces pombe, S. pombe）972h-及prp2-1菌株的新生RNA进行测序：实验先在26℃下培养细胞，随后将其转移至37℃培养2小时，以此获取单转录本的剪接信息。新生RNA从酵母染色质组分中提取（Carrillo Oesterreich、Preibisch、Neugebauer，《Molecular Cell》，2010）；通过连接反应将3' DNA接头标记至新生RNA的3'末端，该接头序列可作为全长反转录的模板，随后通过转录组范围的PCR扩增获取双链cDNA。本数据集包含每个菌株的3次生物学重复数据。样本解复用的脚本可在我的GitHub账号（https://github.com/lherzel）中获取。