Notes: The authors characterized mouse neocortical interneurons that express 5-HT_3A, a ligand-gated cation channel activated by 5-hydroxytryptamine (serotonin), during embryonic development. Transgenic mice that expressed green fluorescent protein (GFP) under the control of the 5-HT_3A promoter were created. Single 5-HT_3A-expressing neurons within 300µm brain sections of transgenic mice at various stages of embryonic development were subjected to whole-cell path-clamp recordings to examine their electrophysiological properties. To confirm activation of the 5-HT_3A promoter in these cells, GFP expression was visualized by fluorescence microscopy without breaking the patch clamp seal. The contents of these single neurons then were aspirated and expelled into a 10µl reverse transcription reaction. After the reverse transcription, PCR was performed to simultaneously detect mRNAs encoding two isoforms of glutamic acid decarboxylase, three calcium-binding proteins, three neuropeptides, two transcription factors and reelin, a protein thought to be involved in neuronal migration and morphology. Two rounds of PCR using nested primers were required to detect these mRNAs. PCRs were performed using GoTaq® DNA Polymerase. Amplified products were visualized by agarose gel electrophoresis, using the 100bp DNA Ladder as a size standard. (4096)

Notes: In this paper, the 100bp DNA Ladder was used on 1.5% agarose gel to determine the size of PCR products generated with Taq DNA Polymerase. (2229)

Notes: Plasmid DNA was purified with the Wizard® Plus Miniprep DNA Purification System and further purified for microinjection. Transgenic zebrafish were identified by PCR using Taq DNA Polymerase. PCR products were resolved on gels with the 100bp DNA Ladder as a size marker. (1183)