Multiplex qPCR of Single Cells

cDNA synthesis was performed using the Single Cell-to-CT Kit (Invitrogen), and multiplex qPCR was performed using the TaqMan Gene Expression Assay system (Applied Biosystems). All TaqMan probes were purchased from Applied Biosystems and are as follows: Arpp21 (Mm00473630_m1), Cck (Mm00446170_m1), Chat (Mm01221880_m1), Chodl (Mm00507273_m1), Crhbp (Mm01283832_m1), Dlx1 (Mm00438424_m1), Gad1 (Mm04207432_g1), Gad2 (Mm00484623_m1), Gal (Mm00439056_m1), Hcrtr1 (Mm01185776_m1), Hcrtr2 (Mm01179312_m1), Htr3a (Mm00442874_m1), Lhx6 (Mm01333348_m1), Meis2 (Mm00487748_m1), Noct (Mm00802276_m1), Nos1 (Mm01208059_m1), Npy (Mm01410146_m1), Nr2f2 (Mm00772789_m1), Nrgn (Mm01178296_g1), Nts (Mm00481140_m1), Pmch (Mm01242886_g1), Pou3f3 (Mm00843792_s1), Pvalb (Mm00443100_m1), Reln (Mm00465200_m1), Snord116 (Mm05911478_g1), Sst (Mm00436671_m1), Stim2 (Mm01223103_m1), Tac1 (Mm01166996_m1), Tac2 (Mm01160362_m1), Vglut1 (Mm00812886_m1), Vglut2 (Mm00499876_m1). Target amplification was performed using the CFX Opus Real-Time PCR System (384 well, Bio-Rad) with Bio-Rad CF Maestro 1.1 software version 4.1.

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Tossell K., Yu X., Giannos P., Anuncibay Soto B., Nollet M., Yustos R., Miracca G., Vicente M., Miao A., Hsieh B., Ma Y., Vyssotski A.L., Constandinou T., Franks N.P, & Wisden W. (2023). Somatostatin neurons in prefrontal cortex initiate sleep-preparatory behavior and sleep via the preoptic and lateral hypothalamus. Nature Neuroscience, 26(10), 1805-1819.

Publication 2023

Single Cell-to-CT Kit TaqMan Gene Expression Assay system TaqMan probes CFX Opus Real-Time PCR System

Assay Cdna Cell Gad1 Gene expression Htr3a Nos1 Reln Synthesis

Corresponding Organization :

Other organizations : Imperial College London, Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, UK Dementia Research Institute, ETH Zurich, SIB Swiss Institute of Bioinformatics, University of Zurich

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Variable analysis

independent variables

CDNA synthesis kit used (Single Cell-to-CT Kit, Invitrogen)
Multiplex qPCR assay system used (TaqMan Gene Expression Assay, Applied Biosystems)
TaqMan probes used (list provided)

dependent variables

Expression levels of the following genes: Arpp21, Cck, Chat, Chodl, Crhbp, Dlx1, Gad1, Gad2, Gal, Hcrtr1, Hcrtr2, Htr3a, Lhx6, Meis2, Noct, Nos1, Npy, Nr2f2, Nrgn, Nts, Pmch, Pou3f3, Pvalb, Reln, Snord116, Sst, Stim2, Tac1, Tac2, Vglut1, Vglut2

control variables

Mice/samples used for gene expression analysis (not explicitly mentioned)

controls

No positive or negative controls were explicitly mentioned in the input.

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