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Powerplex 16 str system

Manufactured by Promega
Sourced in United States

The PowerPlex® 16 STR system is a DNA profiling kit developed by Promega. It is designed to amplify short tandem repeat (STR) loci for human identification and relationship testing purposes. The system allows for the simultaneous amplification of 16 STR loci in a single reaction.

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8 protocols using powerplex 16 str system

1

DNA Profiling via STR Analysis

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DNA samples at passages 25, 34 and 44 were amplified by the PowerPlex® 16 STR System (Promega, Tokyo, Japan) and repeat numbers were determined by the ABI 3500 Genetic Analyzer.
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2

HeLa Cell Culture Protocol

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The human cervix adenocarcinoma HeLa cell line was obtained from American Type Culture Collection and was authenticated with the Promega PowerPlex® 16 STR system (Madison, WI) in October 2012. HeLa cells were grown in DMEM supplemented with 10% heat-inactivated FBS and 1% penicillin and streptomycin. Cells were maintained in a humidified atmosphere containing 5% CO2 at 37°C.
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3

STR Profiling for Genetic Analysis

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The DNA sample was amplified by the PowerPlex 16 STR System (Promega) and repeat numbers were determined by the ABI 3500 Genetic Analyzer.
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4

Induction of FOXM1 in TE-1 Esophageal Cancer Cells

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The ESCC-derived cell line, TE-1, was kindly donated by Dr. Pierre Hainaut (University of Grenoble, France). TE-1 cells were cultured in RPMI medium (Invitrogen) supplemented with 10% fetal bovine serum (Thermo) and 1% of the cocktail penicillin/glutamine/streptomycin (Invitrogen) and maintained at 37 °C under 10% CO2. TE-1 cells had FOXM1 expression levels induced by transfecting different amounts of an expression vector (pcDNA3-FOXM1) [12 (link)] or the empty backbone vector (pcDNA3, Invitrogen), used as an experimental control, using Lipofectamine 2000 (Invitrogen), following the manufacturer’s protocol. Cells were authenticated using Powerplex 16 STR System (Promega, Madison, WI, USA) and were routinely tested for mycoplasma using a Mycosensor (Agilent, Santa Clara, CA, USA).
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5

Establishing Crizotinib-Resistant Cell Lines

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HCC78 cells harboring the SLC34A2‐ROS1 fusion gene were kindly provided by Dr William Pao (Vanderbilt University, Nashville, TN, USA). ABC‐20 cells were established in our laboratory from pleural effusion obtained from a Japanese male former smoker who had lung adenocarcinoma harboring the CD74‐ROS1 fusion gene. The experiment regarding ABC‐20 cells was approved by the Institutional Review Board of Okayama University Hospital. Written informed consent was obtained from the patient. PC‐9 cells harboring EGFR 19 del E746_A750 were purchased from the European Collection of Cell Cultures (Salisbury, UK). 293T cells were purchased from the RIKEN Cell Bank (Ibaragi, Japan). Cells were cultured in RPMI 1640 medium (Sigma‐Aldrich, St. Louis, MO, USA) supplemented with 10% heat‐inactivated FBS and 1% penicillin/streptomycin in a tissue culture incubator at 37°C with 5% CO2. To establish a crizotinib‐resistant cell line, HCC78 cells were treated with gradually increasing concentrations of crizotinib, starting at .2 μmol/L (lower than the IC50 of HCC78 cells). After 4 months, the cells grew in the presence of 2 μmol/L crizotinib and were designated as HCC78R cells. HCC78R cells were maintained in culture medium containing 1 μmol/L crizotinib. The resistant cell lines were tested using the PowerPlex 16 STR System (Promega Corporation, Madison, WI, USA).
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6

Comprehensive Cell Line Characterization

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The human NSCLC cell lines RERF-LC-AI, RERF-LC-KJ, and LC2/Ad were obtained from Riken BRC through the National Bio-Resource Project of the MEXT (Tsukuba, Japan). The A549 cells were purchased from both Riken BRC and American Type Culture Collection (Manassas, VA), while the PC–9 cell line was obtained from IBL cell bank (Gunma, Japan). The genotypes of all cell lines were identified with STR Identifier (Applied Biosystems) or PowerPlex 16 STR system (Promega). All the cell lines were maintained in culture in RPMI 1640 medium with 2mM L-glutamine (Invitrogen) supplemented with 10% FBS (Sigma-Aldrich) (A549, RERF-LC-AI, RERF-LC-KJ and PC–9) or 15% FBS (LC2/Ad) and 50 U/ml penicillin streptomycin (Sigma-Aldrich) at 37°C in a humidified atmosphere with 5% CO2. For cell culture work, Gefitinib (Cayman, #13166), Docetaxel (Enzo Life Sciences, BML-T129), LY294002 (Cayman, #70920), PD98059 (Cayman, #10006726) and KU55933 (Tocris Bioscience, #3544) were dissolved in DMSO (Sigma-Aldrich). Gemcitabine (Tocris Bioscience, #3259), Pemetrexed (Santa Cruz, #sc–219564), Vinorelbine (Santa Cruz, #sc–216059), Caffeine (Tocris Bioscience, #2793) and N-acetyl-L-cysteine (NAC) (Sigma-Aldrich, #A9165) were dissolved in PBS (-).
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7

NSCLC Cell Lines Characterization and Culture

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The human NSCLC cell lines LC‐2/ad, A549, RERF‐LC‐AI, and RERF‐LC‐KJ were obtained from Riken BRC through the National Bio‐Resource Project of the MEXT (Tsukuba), while PC‐9 was obtained from the IBL cell bank (Gunma). The genotypes of all cell lines were identified using the PowerPlex 16 STR system (Promega). The cell lines were maintained as previously described.9 For cell culture, tofacitinib (#S5001, Selleck), gefitinib (#13166; Cayman), LY294002 (#70920; Cayman), PD98059 (#10006726; Cayman), and PF4708671 (#4032; Tocris) stock solutions were prepared in DMSO (Sigma‐Aldrich), whereas recombinant human IFNγ (#11500; PBL Assay Science) and epidermal growth factor (EGF) (#236‐EG; R&D Systems) stock solutions were prepared in PBS (−).
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8

HeLa Cell Line Authentication and Culture

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The human cervix adenocarcinoma cell line HeLa was obtained from American Type Culture Collection and was authenticated with the Promega PowerPlex® 16 STR system in October 2012. HeLa cells were maintained in DMEM supplemented with 10% heat-inactivated FBS, 1% penicillin and streptomycin in a humidified atmosphere containing 5% CO 2 at 37°C.
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