Transcriptome profiling of sorted endoreduplicated nuclei from mature-green tomato fruits. TomEndoRNA-Seq

In this study, we have tested the feasibility of a RNA-Seq approach using total nuclear RNA extracted from purified populations of flow cytometry-sorted nuclei based on their DNA content. Nuclei populations from tomato mature-green fruit pericarp tissue were sorted by flow cytometry according to their DNA content. Total nuclear RNA were then extracted. A total of 24 cDNA libraries were then constructed corresponding to 4 ploidy levels (4C, 8C, 16C and 32C) in 3 replicates, with duplex-specific nuclease (DSN) or without (no-DSN) treatment. The multiplex sequencing reaction was performed on an Illumina HiSeq 2000 machine.

本研究针对基于DNA含量的流式细胞术分选纯化细胞核群体，提取总核RNA并验证RNA测序（RNA-Seq）方法的可行性。本研究以番茄绿熟期果实果皮组织的细胞核群体为材料，依据其DNA含量通过流式细胞术完成分选，随后提取所得细胞核的总核RNA。最终构建24个cDNA文库，覆盖4个倍性水平（4C、8C、16C与32C），每个倍性水平设置3次生物学重复，并分别开展双链特异性核酸酶（duplex-specific nuclease, DSN）处理与未处理（无-DSN）两种分组。所有文库采用Illumina HiSeq 2000测序平台完成多重测序反应。