Gibco™ DMEM, high glucose

Mammalian cell culture media MDA-MB-231

Experiment
Mammalian cell culture media MDA-MB-231
Product
Gibco™ DMEM, high glucose from Fisher Scientific
Manufacturer
Fisher Scientific

Protocol tips

Protocol tips
supplemented with 10% fetal bovine serum.

Publication protocol

SK-BR-3, MCF-7, and MDA-MB-231 human breast cancer cell lines as well as the MCF-10A human normal mammary epithelial cell line were obtained from the American Type Culture Collection. All cell lines were cultured at 37 °C in a humid atmosphere with 5% CO2. MCF-7 and SK-BR-3 cells were cultured in RPMI-1640 (Gibco, USA) supplemented with 10% fetal bovine serum (Gibco, USA). MDA-MB-231 cells were cultured in Dulbecco's modified Eagle's medium (Gibco, USA) supplemented with 10% fetal bovine serum. MCF-10A cells were cultured in MEGM (Lonza, Switzerland) supplemented with 100 ng/mL cholera toxin (Sigma, USA). The Cell-SELEX library and primers were purchased from Sangon Biotech Co., Ltd. (Shanghai, China). The library contained a randomized region of 40 nt flanked by two constant regions of 23 nt on both sides for primer annealing and PCR amplification (5′-AGCAGAGTTCACGACCCGATAAG-N40-GAGTTACATACCAATCGTCGCAG-3′). For PCR amplification, the forward primer was labeled with FAM at the 5′ end (5′-FAM-AGCAGAGTTCACGACCCGATAAG-3′) to monitor the enrichment of the library pool by flow cytometry (ACEA NovoCyteTM Series, ACEA, USA), and the reverse primer was labeled with biotin at the 5′ end (5′-biotin-CTGCGACGATTGGTATGTAACTC-3′) to isolate the sense strands using streptavidin-coated magnetic nanoparticles (SA@MNPs) for subsequent selection rounds. The SA@MNPs were synthesized according to our previous report 45. The agarose used for electrophoretic analysis was purchased from Sangon Biotech Co., Ltd. (Shanghai, China). The binding buffer utilized in the Cell-SELEX process was prepared with phosphate-buffered saline (PBS, pH 7.4) supplemented with 4.5 g/L glucose, 5 mM MgCl2, 0.1 mg/mL yeast tRNA, and 1 mg/mL bovine serum albumin. The washing buffer was prepared with PBS supplemented with 4.5 g/L glucose and 5 mM MgCl2.

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Manufacturer protocol

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