AmpFLSTR™ Identifiler™ PCR Amplification Kit

Cell line authentication Ovarian cancer cell line PA-1

Experiment
Cell line authentication Ovarian cancer cell line PA-1
Product
AmpFLSTR™ Identifiler™ PCR Amplification Kit from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Protocol tips
Compare profiles to consolidated (ATCC, DSMZ, JCRB and RIKEN) databases

Publication protocol

DNA isolation and STR profiling: Cell lines were grown in appropriate specific standard media. Genomic DNA was isolated from 0.5–5 × 106 cells using a Zymo Research ZR genomic DNA II kit and quantified by gel electrophoresis and ethidium bromide staining by comparison to a DNA mass ladder. Multiplex PCR amplified products were generated using 1–2 ng of genomic DNA with an Applied Biosystems Identifiler kit and ABI 3730 capillary sequencer as described [2, 18]. STR loci were analyzed with Gene Mapper 4.0. Profiles were compared to published reports [22, 25], consolidated (ATCC, DSMZ, JCRB and RIKEN) databases, and an in-house database, using a custom search algorithm designed to facilitate comparison of cell lines with related profiles and identify individual cell lines in a mixture (C. Korch and J. West, Vanderbilt University, unpublished). STR profiles of the ovarian and endometrial cancer cells analyzed in this study are available online at http://DNAsequencingcore.UCDenver.edu.

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Manufacturer protocol

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