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An automated approach to prepare tissue-derived spatially barcoded RNA-sequencing libraries

机译:制备组织衍生的空间条形码RNA测序文库的自动化方法

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摘要

Sequencing the nucleic acid content of individual cells or specific biological samples is becoming increasingly common. This drives the need for robust, scalable and automated library preparation protocols. Furthermore, an increased understanding of tissue heterogeneity has lead to the development of several unique sequencing protocols that aim to retain or infer spatial context. In this study, a protocol for retaining spatial information of transcripts has been adapted to run on a robotic workstation. The method spatial transcriptomics is evaluated in terms of robustness and variability through the preparation of reference RNA, as well as through preparation and sequencing of six replicate sections of a gingival tissue biopsy from a patient with periodontitis. The results are reduced technical variability between replicates and a higher throughput, processing four times more samples with less than a third of the hands on time, compared to the standard protocol.
机译:测序单个细胞或特异性生物样品的核酸含量越来越常见。这驱动了需要强大,可扩展和自动库准备协议的需求。此外,增加对组织异质性的理解已经导致旨在保留或推断空间背景的几种独特测序方案的发展。在本研究中,用于保留转录物的空间信息的协议已经适于在机器人工作站上运行。通过制备参考RNA的鲁棒性和可变性来评估方法空间转录组科,以及通过从牙周炎的患者的患者的牙龈组织活检的六种复制部分的制备和测序来评估。结果减少了复制与较高吞吐量之间的技术可变性,与标准协议相比,处理较少的样品的比例少于三分之一的样本。

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