title: GSM8121068: DSCL cases_2; Homo sapiens; RNA-Seq
library_construction_protocol: CD68+ microglia were isolated from white matter groups (n=4 cases per group) from the Cognitive Function and Ageing Study neuropathology cohort by immuno-laser capture microdissection and total RNA was extracted from the LCM-isolated cells using the PicoPure RNA isolation kit (Life technologies: Applied Biosystems, USA). 8-μl of extracted total RNA was reversed transcribed to cDNA and amplified following 20 cycles of amplifying PCR (initial denaturation at 98°C for 45 s, the product was amplified at 98°C for 10 s, 62°C for 15 s, 72°C for 3 min, and then 72°C for 5 min). Following fragmentation, samples underwent adapter ligation and indexing to create cDNA libraries, permitting recognition of each sample once pooled. Each cDNA library product underwent amplification following 9 cycles of amplifying PCR (initial denaturation at 98°C for 30 s, then the product was amplified at 98°C for 10 s, 65°C for 75 s, and then 65°C for 5 min) and pooled. Pooled cDNA libraries were ran in duplicate lanes of a flow cell to maximize the number of mapped reads (100bp single read) and sequenced on an Illumina HiSeq 2500 platform using the Illumina TruSeq SBS Kit CA, United states).