AmpFLSTR™ Identifiler™ PCR Amplification Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Extract DNA from cell pellets using QiaAMP mini preps (Qiagen cat 51306)	Use of a DNA amount of 0.15 ng, so that lower-level mixing of cell lines could be visible Compare the STR profiles to known ATCC fingerprints (ATCC.org), to the Cell Line Integrated Molecular Authentication database

Upstream tips
Extract DNA from cell pellets using QiaAMP mini preps (Qiagen cat 51306)
Protocol tips
Use of a DNA amount of 0.15 ng, so that lower-level mixing of cell lines could be visible Compare the STR profiles to known ATCC fingerprints (ATCC.org), to the Cell Line Integrated Molecular Authentication database

Publication protocol

Cell Line authentication: Cell lines were grown to 70–80% confluence in a T-75 flask, trypsinized and the cell pellets were washed once with 1X phosphate buffered saline. DNA was extracted from cell pellets using QiaAMP mini preps (Qiagen cat 51306) and DNA was quantitated by Nanodrop spectrometry. Cell lines were validated by STR DNA fingerprinting using the AmpFℓSTR Identifiler kit according to manufacturer instructions (Applied Biosystems cat 4322288). A higher DNA amount of 0.15 ng DNA was used so that lower level mixing of cell lines could be visible. The samples were run on an Applied Biosystems 96-well 3730 Genetic Analyzer. Data were analyzed using GeneMapper (Applied Biosystems) and exported for STR comparisons. For the purposes of this paper, we did not change any of the automated calls. The STR profiles were compared to known ATCC fingerprints (ATCC.org), to the Cell Line Integrated Molecular Authentication database (CLIMA) version 0.1.200808 (http://bioinformatics.istge.it/clima/),18 to the complete 16 loci reference listed in Lorenzi, et al.,19 and to the MD Anderson fingerprint database. The STR profiles matched known DNA fingerprints (see Table 1 and Supplemental Table 1). All cell lines were also tested by Giemsa-banding and silver staining of metaphase chromosomes to verify that the cell lines were not cross-contaminated.

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 Renal carcinoma cell line Caki-1 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 Renal carcinoma cell line Caki-1 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.