Profiler plus kit

Manufactured by Thermo Fisher Scientific

The Profiler Plus Kit is a multiplex STR analysis system designed for human identification. It allows for the simultaneous amplification and detection of up to 24 genetic markers in a single reaction. The kit is compatible with a range of instrumentation and software for DNA profiling and analysis.

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Lab products found in correlation

Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Hek293t cell, by American Type Culture Collection (1 mentions) Sodium pyruvate, by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Avantor (1 mentions) Glutamine plus, by Atlanta Biologicals (1 mentions) Glutamax, by Thermo Fisher Scientific (1 mentions) Umuc3, by American Type Culture Collection (1 mentions) Sk mel24, by Thermo Fisher Scientific (1 mentions) Immunomagnetic beads, by STEMCELL (1 mentions) Mycoalert, by Cambrex (1 mentions)

Close spelling variants of this product

AmpFLSTR Profiler Plus PCR Amplification Kit (9 citations) AmpFLSTR-Profiler Plus kit (7 citations) Profiler Plus (6 citations) Profiler Plus PCR Amplification Kit (4 citations) PLUSTM (3 citations) AmpFISTR Profiler Plus kit (2 citations)

6 protocols using profiler plus kit

Cell Line Characterization and Authentication

Cited 2 times

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Bladder cancer cell lines UMUC3, TCCSUP, RT4, CRL2742, SW780, MGHU4, and UMUC9 were previously described (4 (link)). Lung and prostate cancer cell lines, A549 (16 (link)) and 22Rv1 (17 (link)) were obtained from the cell line repository at University of Colorado Cancer Center PPSR. Breast cancer cell lines HCC1806 and BT549 (18 (link)) were a gift from Dr. Richer at the University of Colorado. HEK293T cells were purchased from ATCC. All reagents and media were purchased from Invitrogen. All lines were authenticated by the University of Colorado Cancer Center PPSR core using an Applied Biosystems Profiler Plus Kit that analyzed 9 loci (Life Technologies 4303326). Cells were used in experiments within 6 weeks of thawing and were tested to be free of mycoplasma.

Agarwal N., Dancik G.M., Goodspeed A., Costello J.C., Owens C., Duex J.E, & Theodorescu D. (2016). GON4L drives cancer growth through a YY1-androgen receptor-CD24 axis. Cancer research, 76(17), 5175-5185.

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Authenticating Bladder Cancer Cell Lines

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Human bladder cancer cell lines T24, 253J, UMUC3, J82, MGHU3 and HT1197 were authenticated by the University of Colorado PPSR core using an Applied Biosystems Profiler Plus Kit which analyzed 9 loci (Life Technologies 4303326). Cell ampules were resuscitated less than 2 months prior to being used in experiments in this study. Cells were cultured at 37 degrees Celsius, 5% CO₂ in the following media. T24: Dulbecco’s Modified Eagle’s Medium/F12 (DMEM/F12) + 5% Fetal Bovine Serum (FBS), 253J and HT1197: Modified Eagle’s Medium (MEM) + 10% FBS + 0.1mM Nonessential Amino Acids (NEAA) + 1 mM Sodium Pyruvate, UMUC3 and MGHU3: MEM + 10% FBS + 1 mM Sodium Pyruvate, J82: MEM + 10% FBS + 0.1 mM NEAA. All reagents for media components were obtained from Invitrogen.

Hensel J., Duex J.E., Owens C., Dancik G.M., Edwards M.G., Frierson H.F, & Theodorescu D. (2015). Patient mutation directed shRNA screen uncovers novel bladder tumor growth suppressors. Molecular cancer research : MCR, 13(9), 1306-1315.

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Cell Line Authentication and Culturing

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HCC lines and UMUC3 and T24 were obtained from American Type Culture Collection. UMUC3 and T24 were grown as described (Guin et al. 2014 (link)) except as noted below and were authenticated by the University of Colorado Cancer Center Protein Production Shared Resource using an Applied Biosystems Profiler Plus kit, which analyzed nine STR loci (Life Technologies, 4303326). After authentication, cells were frozen within 1–2 wk. Vials of cells were resuscitated <2 mo prior to being used in experiments in this study. All cells were cultured in DMEM (VWR Scientific) with GlutaminePlus (Atlanta Biologicals), 10% FBS (Seradigm), penicillin/streptomycin (GIBCO), glutamax (GIBCO), and sodium pyruvate (GIBCO), except HepG2 cells were cultured in EMEM (American Type Culture Collection) without sodium pyruvate.

Stern J.L., Theodorescu D., Vogelstein B., Papadopoulos N, & Cech T.R. (2015). Mutation of the TERT promoter, switch to active chromatin, and monoallelic TERT expression in multiple cancers. Genes & Development, 29(21), 2219-2224.

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Protocol for Cutaneous Melanoma Cell Line Cultivation

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TAK-733 (Suppl Figure 1) was provided by Takeda Cambridge US, Inc. (Cambridge, MA) under a materials transfer agreement (MTA) and prepared as a 10mM stock solution in dimethyl sulfoxide (DMSO). For in vivo studies, this compound was prepared as a suspension in 0.5% methylcellulose in sterile water by brief vortexing followed by sonication for 10 minutes. The cutaneous melanoma cell lines were obtained from the American Type Culture Collection (ATCC; Rockville, MD) and were cultured according to their recommendations. All cell lines except SK-MEL24 and Hs852t were maintained in DMEM supplemented with 10% fetal bovine serum, 1% nonessential amino acids, 1% penicillin/streptomycin (Invitrogen; Carlsbad, CA) and were maintained at 37°C under an atmosphere of 95% O2, 5% CO2. SK-MEL 24 required 15% fetal bovine serum and Hs852t required a 10% CO2 atmosphere. Authenticity of all cell lines was verified by the University of Colorado Cancer Center DNA Sequencing and Analysis Core using the Profiler Plus Kit (Applied Biosystems; Foster City, CA). The data obtained was compared with ATCC data to ensure the cell line profiles had not changed with the latest profiling performed in January and February 2013.

Micel L.N., Tentler J.J., Tan A.C., Selby H.M., Brunkow K.L., Robertson K.M., Davis S.L., Klauck P.J., Pitts T.M., Gangolli E., Fabrey R., O'Connell S.M., Vincent P.W, & Eckhardt S.G. (2014). Antitumor Activity of the MEK Inhibitor TAK-733 Against Melanoma Cell Lines and Patient-Derived Tumor Explants. Molecular cancer therapeutics, 14(2), 317-325.

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Chimerism Determination Post-Transplantation

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Post-transplantation D30 (range, 20-50) and 100 (range, 80-120) chimerism was determined using recipient PB or BM samples collected in EDTA. “Total donor cell” chimerism was performed on buffy coat leukocytes. “T-cell” chimerism was on Ficoll Hypaque separated lymphocytes from which purified CD3+ T-cells were isolated using immunomagnetic beads (Stem Cell Technologies Inc.). Pretransplantation PB samples were used to determine the donor and recipient genotypes based on 9 CODIS Short Tandem Repeat loci using the Applied Biosystems Profiler Plus® kit with the ABI 3130 capillary genetic analyzer to determine the alleles at each locus. Informative alleles unique to either donor or recipient were used to calculate % donor chimerism at each locus using the median peak intensity of amplicons attributable to donor divided by the sum of all amplicons at that locus. In cases where there were only unique recipient amplicons, % donor chimerism was calculated as (100 – % recipient chimerism). 95% confidence interval for chimerism was ±5%.

Koreth J., Kim H.T., Nikiforow S., Milford E.L., Armand P., Cutler C., Glotzbecker B., Ho V.T., Antin J.H., Soiffer R.J., Ritz J, & Alyea EP I.I.I. (2014). Donor Chimerism Early after Reduced-intensity Conditioning Hematopoietic Stem Cell Transplantation Predicts Relapse and Survival. Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation, 20(10), 1516-1521.

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Cell Line Authentication and Maintenance

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The CRC cell lines examined in this study were purchased from the American Type Culture Collection (ATCC), European Collection of Cell Cultures (ECACC) and Korean Cell Line Bank (KCLB)—Seoul National University. Cells were maintained and treated in RPMI media that contained 10% fetal bovine serum, 1% nonessential amino acids, and 1% penicillin/streptomycin. The cells were placed in an incubator at 37°C containing 5% CO₂. The cells were routinely screened for the presence of Mycoplasma (MycoAlert; Cambrex BioScience). The CRC cell lines were treated with dasatinib when they reached ∼70% confluence. All cell lines were tested and authenticated in the University of Colorado Cancer Center DNA Sequencing and Analysis Core. CRC cell line DNA was tested using the Profiler Plus kit (Applied Biosystems). The data obtained were compared with cell line databases to ensure the cell lines have not changed.

Scott A.J., Song E.K., Bagby S., Purkey A., McCarter M., Gajdos C., Quackenbush K.S., Cross B., Pitts T.M., Tan A.C., Eckhardt S.G., Fenton H., Arcaroli J, & Messersmith W.A. (2017). Evaluation of the efficacy of dasatinib, a Src/Abl inhibitor, in colorectal cancer cell lines and explant mouse model. PLoS ONE, 12(11), e0187173.

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