Brain tumor cell line authentication, an efficient alternative to capillary electrophoresis by using a microfluidics-based system

Neuro-oncology
Qian AnGeoffrey J Pilkington

Abstract

The current method for cell line authentication is genotyping based on short tandem repeat (STR)-PCR involving coamplification of a panel of STR loci by multiplex PCR and downstream fragment length analysis (FLA), usually performed by capillary electrophoresis. FLA by capillary electrophoresis is time-consuming and can be expensive, as the facilities are generally not accessible for many research laboratories. In the present study, a microfluidic electrophoresis system, the Agilent 2100 Bioanalyzer, was used to analyze the STR-PCR fragments from 10 human genomic loci of a number of human cell lines, including 6 gliomas, 1 astrocyte, 1 primary lung cancer, 1 lung brain metastatic cancer, and 1 rhabdomyosarcoma; and this was compared with the standard method, that is, capillary electrophoresis, using the Applied Biosystems 3130xl Genetic Analyzer. The microfluidic electrophoresis method produced highly reproducible results with good sensitivity in sizing of multiple PCR fragments, and each cell line demonstrated a unique DNA profile. Furthermore, DNA fingerprinting of samples from 5 different passage numbers of the same cell line showed excellent reproducibility when FLA was performed with the Bioanalyzer, indicating that no cros...Continue Reading

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Citations

Jan 19, 2016·Tumour Biology : the Journal of the International Society for Oncodevelopmental Biology and Medicine·Emelia A AssarSassan Hafizi
Sep 9, 2017·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Zaynah MaherallyGeoffrey J Pilkington
Jan 10, 2020·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Diana M LeiteGeoffrey J Pilkington
Jul 28, 2016·Micromachines·Navid KashaninejadBahar Firoozabadi

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