Instrument: Illumina HiSeq 4000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Dorso-lateral striatum (bregma, AP: 1.42 to -0.58 mm) from Lhx6Cre:RFP, Pvcre:TdT and Htr3aEGFP mice was dissociated into a single cell suspension as described in Zeisel et al., Science vol. 347 p.1138 (2015) . Dissociated cells were FACS sorted based on fluorescence (RFP+ and EGFP+) and positive cells were sorted using BD FACSAria II SORP into a 9600 multiwell array (2400 cells/plate) prefilled with 50 nl lysis buffer (500nM STRT-P1-T31, 4.5nM dNTP, 2% Triton-X-100, 20mM DTT, 1.5U/μl TaKaRa RNase Inhibitor), followed by 3min lysis at 72°C. 85nl reverse transcription (RT) mix (2.1X SuperScript II First-Strand Buffer, 12.6mM MgCl2, 1.79M betaine, 14.7U/μl SuperScript II, 1.58U/μl TaKaRa RNase Inhibitor, 10.5μM P1B-UMI-RNA-TSO) were dispensed and RT carried out 42°C for 90 minutes. 96 Transposome stocks were assembled (6.25μM barcoded adapter (STRT-Tn5-Idx[1-96]), 6.25μM Tn5 transposase, 40% glycerol), 37°C 1h. Tn5 reactions were assembled with 3μl transposome and 2μl amplified cDNA, in a total 20μl 1x CutSmart buffer (NEB), and incubated at 55°C 20min.