A comprehensive RNA handling and transcriptomics guide for high-throughput processing of Plasmodium blood-stage samples.. A comprehensive RNA handling and transcriptomics guide for high-throughput processing of Plasmodium blood-stage samples.

Here we have designed a full processing pipeline of P. falciparum RNA that innclues RNA preservation, extraction, cDNA synthesis and amplification, globin depletion and final transcriptome readout. To validate the protocol we have analyzed transcriptomes of P. falciaprum samples derived from filed studies as well as 3D7 lab strains. This dataset have been used to establish the best conditions for whole transcriptome amplification protocol. Additionally it inclues the validation of bead extraction protocol. It also contains field samples used for validation of the entire protocol. Overall design: Samples were extracted using high-throughput techniques adied by automatics liquid handlers, cDNA was amplified and resulted transcriptomes were analyzed and validated using microarray platform.

本研究设计了一套完整的恶性疟原虫（P. falciparum）RNA处理流程，涵盖RNA保存、提取、互补DNA（cDNA）合成与扩增、珠蛋白去除以及最终转录组检测环节。为验证该实验方案，我们对来源于野外研究以及3D7实验室菌株的恶性疟原虫样本的转录组进行了分析。本数据集用于确立全转录组扩增方案的最优条件，同时包含磁珠提取方案的验证内容，亦纳入了用于验证整套实验流程的野外样本。整体实验设计：样本采用自动化液体处理工作站辅助的高通量技术完成提取，经cDNA扩增后，通过微阵列平台对所得转录组进行分析与验证。