RNA from mouse colonic tissue was isolated using the PureLink RNA Mini Kit (Invitrogen, Carlsbad, CA) and from mouse DRG using the PureLink RNA Micro Kit (Invitrogen) according to the manufacturer's instructions without modifications as described previously.13 (link),15 ,18 (link) All samples underwent an on-column DNase treatment with the PureLink DNase Set (Invitrogen). The elution volumes were 60 µL for mouse colonic tissue RNA and 20 μL for mouse DRG RNA. RNA was aliquoted and stored at −80°C until use. RNA quality was assessed using the 2100 Bioanalyzer Instrument (Agilent Technologies, Inc, Santa Clara, CA) or the 2200 TapeStation System (Agilent Technologies, Inc). All samples had an RNA integrity number of 6.5 or higher.
Protocol detail
RNA Isolation from Mouse Colonic Tissue and DRG
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PureLink RNA Mini Kit
PureLink RNA Micro Kit
PureLink DNase Set
2100 Bioanalyzer Instrument
2200 TapeStation System
Colonic
Dnase
Mouse
Tissue
Corresponding Organization : University of Adelaide
Other organizations : Arena Pharmaceuticals (United States)
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Variable analysis
independent variables
- Tissue type (mouse colonic tissue, mouse dorsal root ganglion)
- RNA quality (assessed using the 2100 Bioanalyzer Instrument or the 2200 TapeStation System)
- RNA isolation protocol (PureLink RNA Mini Kit or PureLink RNA Micro Kit, according to manufacturer's instructions)
- On-column DNase treatment (using the PureLink DNase Set)
- Elution volumes (60 μL for mouse colonic tissue RNA, 20 μL for mouse DRG RNA)
- RNA storage conditions (-80°C)
- None specified
- None specified
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