title: Targeted resequencing of human Glioblastoma Multiforme (GBM) tissues, dependent growth factors Glioblastoma Multiforme stem-like cells (D-GSCs), independent growth factors Glioblastoma Multiforme stem-like cells (I-GSCs) and matched blood
design: Targeted resequencing of human Glioblastoma Multiforme (GBM) tissues, dependent growth factors Glioblastoma Multiforme stem-like cells (D-GSCs), independent growth factors Glioblastoma Multiforme stem-like cells (I-GSCs) and matched blood
library_name: I-GSCs-CL#45_p
library_construction_protocol: Glioblastoma Multiforme (GBM) tissues were collected from patients who underwent surgery. From these Glioblastoma Multiforme stem-like cells (GSCs) were isolated. For Dependent growth factors Glioblastoma Multiforme stem-like cells (D-GSCs), single-cell suspension was plated in NeuroCult NS-A medium containing 20 ng/ml of epidermal growth factor (EGF) and 10 ng/ml of fibroblast growth factor (FGF2) and cultured in humidified atmosphere at 37°C, 5% O2 and 5% CO2. For Independent growth factors Glioblastoma Multiforme stem-like cells (I-GSCs), single cell suspension was plated in NeuroCult NS-A medium without growth factors and cultured in humidified atmosphere at 37°C, 5% O2 and 5% CO2.  Genomic DNA was extracted using the Blood & Cell Culture DNA Midi Kit (Qiagen). Truseq custom amplicon sample kit (Illumina Inc, CA, USA) and Ampliseq kit (Illumina Inc, CA, USA) for targeted capture and library preparation was prepared from 250ng or 20ng of double stranded DNA accordingly to manufacturers instructions.