AmpFLSTR™ Identifiler™ PCR Amplification Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Extract DNA from the cell lines with the QIAamp kit (Qiagen)	Profile for amelogenin and 15 STR microsatellite loci

Upstream tips
Extract DNA from the cell lines with the QIAamp kit (Qiagen)
Protocol tips
Profile for amelogenin and 15 STR microsatellite loci

Publication protocol

DNA Preparation and Short Tandem Repeat (STR) Profiling: To isolate genomic DNA, UACC-SARC1 and MDA-MB-231 cells at passage 20 were trypsinized (0.05% trypsin-EDTA; Gibco/Life Technologies, Grand Island NY), washed with phosphate-buffered saline (PBS), and pelleted by centrifugation. DNA was extracted from the cell lines, whole blood and sarcoma tumor with the QIAamp kit (Qiagen, Valencia, CA). UACC-SARC1 and the parental sarcoma tumor were profiled for amelogenin and 15 STR microsatellite loci using the Identifiler polymerase chain reaction (PCR) kit (Applied Biosystems, Foster City, CA) at the University of Arizona Genetics Core per manufacturer's specifications. PCR products were separated by capillary electrophoresis on an ABI 3730 DNA analyzer. Electropherogram analysis and allelic values assignment were performed using Gene Marker software (Soft Genetics, State College, PA). Heterozygosity (i.e., detection of different alleles) expressed as a percentage was calculated based on any difference in the number of peaks within the 15 STR loci (excluding amelogenin). The fractional allele percentage of similarity was inferred by dividing the number of identical alleles by 32 (the total number of alleles studied) and multiplying the result by 100[12].

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Cell line authentication MDA‐MB‐231 cell line using AmpFLSTR™ Identifiler™ PCR Amplification Kit from Thermo Fisher Scientific.

Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for AmpFLSTR™ Identifiler™ PCR Amplification Kit below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Cell line authentication MDA‐MB‐231 cell line using AmpFLSTR™ Identifiler™ PCR Amplification Kit from Thermo Fisher Scientific. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.