Iso-Seq with 5' cap selection and normalization on adult chicken spleen tissue.. Broiler chicken Iso-Seq Spleen

RNA was extracted from an adult female broiler chicken spleen. 5' cap selection (Teloprime) and normalization (column) was performed on the library. The resulting cDNA was size selected for 0-2kb, 2-4kb, and >4kb parts. Each part was then prepared for Pacbio Iso-Seq sequencing. These were sequenced on an RSII machine. The resulting raw data was converted into bam format then converted into CCS reads using default parameters of the SMRT Package Iso-Seq pipeline. The CCS reads were then filtered for full length non-chimeric reads using the classify module of the Iso-Seq pipeline. The fastq file submitted represents the results from the classify output.

本研究从一只成年雌性肉鸡的脾脏中提取了核糖核酸（RNA）。随后对所构建的文库开展5'端帽子结构筛选（Teloprime）与柱式归一化操作。所得的互补脱氧核糖核酸（cDNA）按照片段长度被分选为0~2kb、2~4kb以及>4kb三个组分。针对每个组分分别制备PacBio Iso-Seq测序文库，所有文库均在RSII测序仪上完成测序。所得原始测序数据首先被转换为二进制比对映射（BAM）格式，随后利用SMRT软件包Iso-Seq分析流程的默认参数，将其转换为环形一致序列（CCS）读段。接着借助Iso-Seq分析流程的classify模块，对CCS读段进行筛选以获取全长非嵌合读段。本次提交的FASTQ文件即为该classify模块输出的分析结果。