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PALM-Seq: simultaneous sequencing of cell-free long RNA and small RNA

By Xi Yang, Wen-Jing Wang, Sujun Zhu, Juan Zeng, Yanru Xing, Qing Zhou, Zhongzhen Liu, Haixiao Chen, Jinghua Sun, Liqiang Li, Jinjin Xu, Chunyu Geng, Xun Xu, Jian Wang, Huanming Yang, Shida Zhu, Fang Chen, Wen-Jing Wang

Posted 05 Jul 2019
bioRxiv DOI: 10.1101/686055

Cell-free RNA, including both long RNA and small RNA, has been considered important for its biological functions and potential clinical usage, but the major challenge is to effectively sequence them at the same time. Here we present PolyAdenylation Ligation Mediated-Seq (PALM-Seq), an integrated sequencing method for cell-free long and small RNA. Through terminal modification and addition of 3 polyadenylation and 5 adaptor, we could get mRNA, long non-coding RNA, microRNA, tRNA, piRNA and other RNAs in a single library. With target RNA depletion, all these RNAs could be sequenced with relatively low depth. Using PALM-Seq, we identified pregnant-related mRNAs, long non-coding RNAs and microRNAs in female plasma. We also applied PALM-Seq to sequence RNA from amniotic fluids, leukocytes and placentas, and could find RNA signatures associated with specific sample type. PALM-Seq provides an integrated, cost-effective and simple method to characterize the landscape of cell-free RNA, and can stimulate further progress in cell-free RNA study and usage.

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