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Proteinase K working solution was added and sections incubated at 37 В°C for 15 min. 10 ОјL of biotin-labeled probe was added. DNA was denatured for 10 min at 95 В°C and hybridization continued for 1 h at 37 В°C. After wash buffer, post-hybridization reagent was added and incubated for 10 min. Antibody blocking buffer was added and incubated for 10 min, followed by rabbit anti-biotin and bio Biotin during pregnancy