GSM8146133: Methyl Capture, EA3, Cancer Associated Fibroblast, Post-5AzaC Treatment (10 Day); Homo sapiens; OTHER
Tissue blocks were evaluated by a pathologist to identify the tumor adjacent stroma area from which gDNA could be extracted. The FFPE RNA/DNA Purification Plus Kit (Cat # 54300, Norgen Biotek Corp) was used to isolate total gDNA from patient specimens. Genomic DNA is fragmented, quantified and assessed for quantity and quality using an Agilent 2100 Bioanalyzer and a Qubit fluorimeter, respectively. Fragmented gDNA is used to capture methylated DNAs (MethylCollectorTM Ultra, Active Motif, Cat # 55005). Methylated fractions are used to construct standard fragment libraries with Bio Scientific's NEXTFLEX Rapid DNA-Seq Kit 2.0. For NGS, all methyl- captured DNA is sequenced at the University of California, Irvine genomic Core facility. Bio Scientific's NEXTFLEX Rapid DNA-Seq