GSM9669413: GrnKO, 8, low; Mus musculus; RNA-Seq
Samples were sorted using a BD Aria III sorter. Lipofuscinosis was assessed by 488-autofluorescence, and Cd11b+/CD45+ cells were sorted into 'high' and 'low' autofluorescence populations. 1,000 cells were sorted per sample into 57.5ul  ClonTech Lysis/RT buffer (Takara; Cat# 635013) with RNase inhibitor (Takara; Cat#2313A) in 1.5mL DNA lo-bind tubes, snap frozen, and stored at -80 oC until use. Samples were processed using the Clontech SMARTSeq v4 3′ DE (Takara Bio USA, Inc. 635040 ) kit for cDNA synthesis and the Nextera XT DNA Library Prep Kit (Illumina FC-131-1096) for library generation. 1000 microglia per sample was sorted directly into lysis/RT buffer, and an aliquot of the lysate was used for reverse transcription followed by cDNA amplification to produce full length cDNA. cDNA was normalized to 300 pg of input for library preparation. The tagmentation step simultaneously fragmented and introduced adaptors to the DNA strands, followed by 15 cycles of PCR amplification. Library quantity and quality were assessed with TapeStation D1000 screentape (Agilent 5067-5582). Libraries were pooled in equimolar ratios for sequencing on one lane of an Illumina NovaSeq X Series 10B flowcell, 100 bp; single end reads were generated.