Publikasjoner
NIBIOs ansatte publiserer flere hundre vitenskapelige artikler og forskningsrapporter hvert år. Her finner du referanser og lenker til publikasjoner og andre forsknings- og formidlingsaktiviteter. Samlingen oppdateres løpende med både nytt og historisk materiale. For mer informasjon om NIBIOs publikasjoner, besøk NIBIOs bibliotek.
2019
Forfattere
Fatima Heinicke Xiangfu Zhong Manuela Zucknick Johannes Breidenbach Arvind Yegambaram Meenakshi Sundaram Siri Tennebø Flåm Magnus Leithaug Marianne Dalland Andrew Farmer Jordana M. Henderson Melanie A. Hussong Pamela Moll Loan Nguyen Amanda McNulty Jonathan M. Shaffer Sabrina Shore Hoichong Karen Yip Jana Vitkovska Simon Rayner Benedicte Alexandra Lie Gregor Duncan GilfillanSammendrag
High-throughput sequencing is increasingly favoured to assay the presence and abundance of microRNAs (miRNAs) in biological samples, even from low RNA amounts, and a number of commercial vendors now offer kits that allow miRNA sequencing from sub-nanogram (ng) inputs. Although biases introduced during library preparation have been documented, the relative performance of current reagent kits has not been investigated in detail. Here, six commercial kits capable of handling <100ng total RNA input were used for library preparation, performed by kit manufactures, on synthetic miRNAs of known quantities and human total RNA samples. We compared the performance of miRNA detection sensitivity, reliability, titration response and the ability to detect differentially expressed miRNAs. In addition, we assessed the use of unique molecular identifiers (UMI) sequence tags in one kit. We observed differences in detection sensitivity and ability to identify differentially expressed miRNAs between the kits, but none were able to detect the full repertoire of synthetic miRNAs. The reliability within the replicates of all kits was good, while larger differences were observed between the kits, although none could accurately quantify the relative levels of the majority of miRNAs. UMI tags, at least within the input ranges tested, offered little advantage to improve data utility. In conclusion, biases in miRNA abundance are heavily influenced by the kit used for library preparation, suggesting that comparisons of datasets prepared by different procedures should be made with caution. This article is intended to assist researchers select the most appropriate kit for their experimental conditions.
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Forfattere
Åge Arild NyborgSammendrag
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Forfattere
Pia Heltoft ThomsenSammendrag
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Forfattere
Pia Heltoft Thomsen Trygve S. Aamlid Karin Juul Hesselsøe Karin Normann Bjarni Hannesson Kemeng XiaoSammendrag
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