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北海道大学水産科学研究彙報 = Bulletin of Fisheries Sciences, Hokkaido University >
第65巻 第3号 >

ニホンウナギ卵の母性mRNA次世代シーケンスのためのrRNA除去法の検討

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Please use this identifier to cite or link to this item:http://doi.org/10.14943/bull.fish.65.3.125

Title: ニホンウナギ卵の母性mRNA次世代シーケンスのためのrRNA除去法の検討
Other Titles: Selective Depletion of Ribosomal RNA from Total RNA for Next-generation Sequencing Analysis of Maternal mRNA in Eggs of the Japanese Eel Anguilla japonica
Authors: 泉, ひかり1 Browse this author
萩原, 聖士2 Browse this author
玄, 浩一郎3 Browse this author →KAKEN DB
井尻, 成保4 Browse this author →KAKEN DB
足立, 伸次5 Browse this author →KAKEN DB
Authors(alt): Izumi, Hikari1
Hagihara, Seishi2
Gen, Koichiro3
Ijiri, Shigeho4
Adachi, Shinji5
Keywords: Maternal mRNA
Egg quality
Ribosomal RNA depletion
Next-generation sequencing
Japanese eel
Fish
Issue Date: 20-Dec-2015
Publisher: 北海道大学大学院水産科学研究院
Journal Title: 北海道大学水産科学研究彙報
Volume: 65
Issue: 3
Start Page: 125
End Page: 130
Abstract: Removal of ribosomal RNAs(rRNA) from total RNA in eggs before preparation of a RNA library is required for the analysis of maternal mRNA using next-generarion sequencing ; this mRNA is essential for normal development. In mammals, the rRNA-depletion method is well-established and is already being used for the development of experiment kits. However, this method is not designed for other species, e.g., non-model organisms. In the current study, we attempted to remove rRNA from total RNA of Japanese eel Anguilla japonica, which is one of the most important aquaculture species in Japan. Removal was performed via hibridization-selection using mammalian probes from a commmercially available kit and/or an ordinally designed eel probe. Quantitative polymerase chain reaction(PCR) was performed to examine the depletion efficiency of 18S rRNA and 28S rRNA were comparative between each method, and >98% depletion was achieved. In addition, RNA-sequencing analysis using a next-generation sequencer and read-mapping analysis were carried out using the rRNA-depleted RNA, which was prepared using a mammalian probe, and mapping analysis revealed that rRNA reads were <1% of the total reads. These results suggest that an rRNA-depletion kit designed for a mammal can also be used for fish species.
Type: bulletin (article)
URI: http://hdl.handle.net/2115/60314
Appears in Collections:北海道大学水産科学研究彙報 = Bulletin of Fisheries Sciences, Hokkaido University > 第65巻 第3号

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