The Genomics Shared Resource provides Human Cell Line Authentication service using short tandem repeats (STR) profiling.
The cell lines are important research tools for basic biomedical and pre-clinical research. However, contamination and misidentification of cell lines is a major problem leading to spurious results and irreproducibility of data. Recent changes in NIH grant submission guidelines require all cell lines to be authenticated by chromosomal analysis or short tandem repeat (STR) profiling.
The cell line authentication is performed using short tandem repeat (STR) loci which are also used in DNA fingerprinting. The STR markers are polymorphic DNA loci that contain repeated nucleotide sequence and the number of repeat varies for each individual, combination of repeats are used to match cell lines with their reported profile (ATCC, DSMZ etc). We use AmpFlSTR Profiler PCR Amplification Kit from Applied Biosystems which amplifies 15 short tandem repeat (STR) loci and the amelogenin gene in one single multiplex PCR reaction. The amplification products are fluorescently labeled and analyzed using the ABI 3130xl sequencer. Each peak in the electropherogram represents an allele and by comparison of the sample data to an allelic ladder, the number of repeat units is determined. This numeric value for each repeat is then compared to cell line STR profile reported in ATCC and DSMZ databases to confirm the cell line.
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