Illumina HiSeq 1500 sequencing; The transcriptional signature of human ovarian carcinoma macrophages is associated with extracellular matrix reorganization and disease progression
The transcriptional signature of human ovarian carcinoma macrophages is associated with extracellular matrix reorganization and disease progression
Mononuclear cells were isolated from ascites or peritoneal lavage fluids by density gradient centrifugation (Lymphocyte Separation Medium 1077; PromoCell) and subsequent enrichment on magnetic CD14 microbeads (Miltenyi Biotech). After Ficoll gradient centrifugation, TAMs were purified by CD14 MACS and lysed in Peqgold (TRIzol-like solution based on phenol and chloroform). RNA was digested with DNase (MN) and purified on a Qiagen Minelute column. Illumina-compatible libraries were made using the Epicentre Scriptseq human/mouse/rat low input Gold kit. Libraries were created using the Illumina Truseq mRNA kit v2 used according to the manufacturer's instructions.
Experimental Factor: 31
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