title: Whole genome sequencing of EYFP+ microglia from female mouse
design: Cell nuclei (1500  5000 nuclei per sample) were treated with 15ul proteinase K (0.4ml/ml) for 2h at 50C followed by heat inactivation at 80C for 30minutes. 2ul of the solution was taken for tagmentation with 1ul Tn5 stock solution and 1.6ul reaction buffer (50mM TAPS, 25mM MgCl2, 40% PEG 8k; final concentration 8% PEG, 5mM MgCl2, 10mM TAPS) and 3.4ml H2O (final volume 8ul), incubated at 55C for 10mins, and then inactivated by adding 2ul 0.2% SDS and incubating at 55C for 10mins. For barcoding PCR, each sample was split into 4 individual reaction, each barcoded with a unique molecular barcode (fastq files for the replicate reactions were merged after sequencing). 1ul of the tagmentated DNA solution was used as the template for each barcoding PCR reaction. The PCR program was 72C /3 min, 95C /30 s, [95C /15 s, 67C /30 s, 72C /1 min] x 15 cycles, 72C /5 min, and then 10C hold. After PCR amplification, the reactions were cleaned-up by SPRI-beads (at 0.9X volume) for size selection and were pooled together according in equal parts according to their DNA concentration (qubit dsDNA quantification assays, ThermoFisher). The pooled library was then cleaned-up one more time by SPRI-beads (at 0.9X volume) and sequenced on an Illumina NextSeq 550 using the high output 75bp kit.