title: AB 5500xl Genetic Analyzer paired end sequencing; DNaseq of human Ewing cell line - CADO-ES1
design: DNaseq of human Ewing cell line - CADO-ES1
library_construction_protocol: Ewing Sarcoma cell lines CADO-ES1, was purchased form DSMZ. Ewing sarcoma cell lines were cultivated in RPMI1640 medium, supplemented with 10% (v/v) FCS, 1% Glutamin and 1% of an antibiotic cocktail containing penicillin and streptomycin. All Ewing sarcoma cell lines were maintained on cell culture plastic coated with collagen (0,5mg/ml). Culture media was supplemented by 1% of an antibiotic cocktail containing penicillin and streptomycin and incubated at 37°C and 5%CO2 in a humidified atmosphere. Routine microbiological analysis was done using a PCR Mycoplasma Detection Kit according to suggested protocols (Applied Biosystems, Darmstadt, Germany). The extraction of nucleic acids from cell cultures was performed by the use of the DNeasy Kit (Qiagen) according to the supplier's recommendations. For DNA library preparation 500 – 5000ng DNA were used. ERCCs as an additional internal quality control were added to the libraries. We followed exactly the Thermo-Fischer/Life-Technology(ABI) protocols for the SOLID-5500XL Technology for DNaseq. Most of the procedures were run in a core lab facility of Life-Technology as a commercial service.
attributes.tag: Experimental Factor: disease
attributes.value: Ewing sarcoma