GSM7844483: microglia, WT, biol rep 1; Homo sapiens; RNA-Seq
RNA was extracted at three time points across 2 separate differentiations. Biological replicates were differentiated at either separte times or in separate 6-well plates. Total RNA was collected using the Rneasy Mini kit (Qiagen). Libraries were prepared by the Van Andel Genomics Core from 10 ng of total RNA using Takara SMARTer Stranded Total RNA-Seq Kit v3 Pico Input Mammalian (Takara Bio USA, Moutain View CA) per the manufacturer's protocol. In brief, RNA was sheared to 300-400 bp, after which dscDNA was generated using a template switching mechanism, and unique dual indexed adapters were added to each sample. Ribosomal cDNA was degraded by scZapR and scrRNA probes, and libraries amplified with 13 cycles of PCR. Quality and quantity of the finished libraries were assessed using a combination of Agilent DNA High Sensitivity chip (Agilent Technologies, Inc.), QuantiFluor® dsDNA System (Promega Corp., Madison, WI, USA), and Kapa Illumina Library Quantification qPCR assays (Kapa Biosystems).  Individually indexed libraries were pooled and 100 bp, paired-end sequencing was performed on an Illumina NovaSeq6000 sequencer (Illumina Inc., San Diego, CA, USA), to return a minimum read depth of 50M read pairs per library.