Computational protocol: Fatty acid synthase affects expression of ErbB receptors in epithelial to mesenchymal transition of breast cancer cells and invasive ductal carcinoma

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Protocol publication

[…] Total RNA was extracted from cells/tissues using Tripure (cat. no. 1667165001; Roche Diagnostics, Basel, Switzerland), and reverse transcription to cDNA was performed using the RevertAid First Strand cDNA Synthesis kit (cat. no. K1621; Fermentas; Thermo Fisher Scientific, Inc.), according to the manufacturer's protocol. Primer sequences () were designed using the online software Primer3 (version 0.4.0; Whitehead Institute, Cambridge, MA, USA). Primer sequence specificity was verified using the Basic Local Alignment Search Tool (blast.ncbi.nlm.nih.gov/Blast.cgi), and the primers were synthesized by Shanghai Bioengineering Co. (Shanghai, China). RT-PCR was performed according to the manufacturer's protocol (cat. no. k0221; Fermentas; Thermo Fisher Scientific, Inc.). The thermocycling conditions were as follows: 95°C for 10 min, followed by 40 cycles of 95°C for 15 sec, 60°C for 30 sec and 72°C for 30 sec. The primer annealing temperature was 60°C for all RT-PCR procedures performed. Data were analyzed according to the 2−ΔΔCq method (), and all experiments were repeated three times. […]

Pipeline specifications

Software tools Primer3, BLASTN
Application qPCR
Organisms Mus musculus
Diseases Breast Neoplasms, Neoplasms, Carcinoma, Ductal
Chemicals Cerulenin