Small RNA-seq was performed as described previously (27 (link)). In brief, small RNAs were enriched from 2 μg of total RNA using the Magnetic bead cleanup module (Life Technologies). Libraries were then prepared using the Ion Total RNA-Seq Kit v2 (Life Technologies) and analysed via BioAnalyzer HS Chip (Agilent CA, USA). Afterwards, libraries (18 pM each) were clonally amplified by emulsion PCR in the IonTorrent OneTouch System (Life Technologies) using the Ion PGM Template OT2 200 Kit (Life Technologies) according to manufacturer′s protocol. Sequencing was performed in an IonTorrent Personal Genome Machine using an IonTorrent 316 Chip (Life Technologies).
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Small RNA Sequencing Protocol
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Corresponding Organization : German Primate Center
Other organizations : Institute of Molecular Bioimaging and Physiology
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Variable analysis
independent variables
- Small RNA-seq performed as described previously (27)
- Expression levels of small RNAs
- 2 μg of total RNA used for small RNA enrichment
- Magnetic bead cleanup module (Life Technologies) used for small RNA enrichment
- Ion Total RNA-Seq Kit v2 (Life Technologies) used for library preparation
- Libraries analyzed via BioAnalyzer HS Chip (Agilent CA, USA)
- Libraries (18 pM each) clonally amplified by emulsion PCR in the IonTorrent OneTouch System (Life Technologies) using the Ion PGM Template OT2 200 Kit (Life Technologies)
- Sequencing performed in an IonTorrent Personal Genome Machine using an IonTorrent 316 Chip (Life Technologies)
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