An automated approach to prepare tissue-derived spatially barcoded RNA-sequencing libraries.
2016 (English)In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 6, 37137Article in journal (Refereed) Published
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.
Place, publisher, year, edition, pages
Nature Publishing Group, 2016. Vol. 6, 37137
IdentifiersURN: urn:nbn:se:kth:diva-196766DOI: 10.1038/srep37137ISI: 000388080900001PubMedID: 27849009ScopusID: 2-s2.0-84995665687OAI: oai:DiVA.org:kth-196766DiVA: diva2:1048438
FunderKnut and Alice Wallenberg FoundationSwedish Foundation for Strategic Research Swedish Research Council
QC 201611242016-11-212016-11-212016-12-14Bibliographically approved