Total RNA was isolated from primary cells or human hippocampal tissue using Direct-zol RNA MicroPrep and Direct-zol RNA Miniprep respectively according to manufacturer’s protocol (Zymo Research Corp, CA, USA). Total RNA concentrations were determined using a spectrophotometer (NanoDrop Lite, ThermoScientific, CA, USA). Purity of samples was assessed with 2100 bioanalyzer (Agilent, CA, USA). cDNA was produced from 1000 ng RNA using NCODE Vilo cDNA synthesis kit (microRNAs) or SuperScript III First-Strand Synthesis kit (mRNAs) following manufacturer’s protocol (Life Technologies, CA, USA). SYBR green detection for RT-PCR detailed protocol described previously [14 (link),17 (link)]. The TargetScan prediction software was utilized to identify microRNAs that had conserved 8mer, 7mer or 6mer target sites on the 3’ UTR of SLCI1A2.
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Profiling miRNA and mRNA in hippocampal samples
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Corresponding Organization : University of California, Irvine
Other organizations : University of California, Merced, University of Queensland
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Variable analysis
independent variables
- Isolation of total RNA from primary cells or human hippocampal tissue using Direct-zol RNA MicroPrep and Direct-zol RNA Miniprep respectively
- Total RNA concentrations determined using a spectrophotometer (NanoDrop Lite)
- Purity of samples assessed with 2100 bioanalyzer
- CDNA produced from 1000 ng RNA using NCODE Vilo cDNA synthesis kit (microRNAs) or SuperScript III First-Strand Synthesis kit (mRNAs)
- SYBR green detection for RT-PCR
- TargetScan prediction software utilized to identify microRNAs that had conserved 8mer, 7mer or 6mer target sites on the 3' UTR of SLCI1A2
- Manufacturer's protocols followed for RNA isolation, cDNA synthesis, and RT-PCR
- Not explicitly mentioned
- Not explicitly mentioned
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