Gene mapper id

The genetic assessment of the present study employed the method reported by Bonassi et al. [75] (link). DNA extraction and genotyping were performed by ACGT, Inc. (Wheeling, IL). DNA was extracted by using Oragene DNA purification reagent and its concentrations were evaluated through spectroscopy (NanoDrop Technologies, USA). Each DNA sample was increased by polymerase chain reaction (PCR) for the OXTR gene rs53576 region target with the primers 5-GCC CAC CAT GCT CTC CAC ATC-3 and 5-GCT GGA CTC AGG AGG AAT AGG GAC-3. A PCR reaction of 20 ll, comprising 1.5 ll of genomic DNA from the test sample, PCR buffer, 1 mM each of the forward and reverse primers, 10 mM deoxyribonucleotides, KapaTaq polymerase, and 50 mM MgCl2 was executed. PCR operation comprised of 15 minute denaturation at 95 ^∘C, and 35 cycles at 94 ^∘C (30 s), 60 ^∘C (60 s), 72 ^∘C (60 s) and a final 10 minute step at 72 ^∘C. PCR reactions were genotyped with an ABI 3730xl Genetic Analyzer (Applied Biosystems Inc.) and normalized with GeneScan 600 LIZ (Applied Biosystems, Inc.) size standards on each sample. Genotypic data were inspected using GeneMapper ID (Applied Biosystems, Inc.).
The same procedure was used to assess the OXTR gene rs2254298 region. However, the forward and reverse primers were instead 5-TGA AAG CAG AGG TTG TGT GGA CAG G-3 and 5-AAC GCC CAC CCC AGT TTC TTC-3 respectively.

Amplimers were analyzed on an automated laser fluorescent ABI PRISM 310 Genetic Analyzer (Thermo Fisher Scientific, Waltham, MA, USA). The electropherograms were generated using the dedicated GeneScan® Analysis and Genotyper® software version 3.7 packages and GeneMapper® ID version 3.2 (Applied Biosystems ® from Thermo Fisher Scientific, Waltham, MA, USA).

ERα-expressing MCF7 breast cancer cells were cultured in Dulbecco’s Modified Eagle Medium DMEM (Gibco, Thermo Scientific, Leicestershire, UK, ref. 41965-239). Media was supplemented with 10% foetal bovine serum (FBS), 50 U/ml penicillin, 50 μg/ml streptomycin and 2 mM L-glutamine. MCF7 cells were obtained from ATCC and they were tested for mycoplasma contamination. Also, the MCF7 cells were genotyped by short-tandem repeat genetic profiling using the PowerPlex_16HS_Cell Line panel and analysed using Applied Biosystems Gene Mapper ID v3.2.1 software by the external provider Genetica DNA Laboratories (LabCorp Specialty Testing Group). For the cell treatments, 4-Hydroxytamoxifen (Sigma-Aldrich, #HG278) or Fulvestrant (Selleckchem, #S1191) were used at final concentration 100 nM.