Genomic mini ax body fluids kit

Peripheral blood (12 ml) was collected into K₃EDTA tubes (Becton–Dickinson, New Jersey, Franklin Lakes, USA). Plasma was separated within an hour by double centrifugation at 300×g and 1000×g, both at 4 °C for 10 min. Aliquots of the plasma were frozen at − 80 °C. DNA was extracted (according to the manufacturer’s instructions) from 1 ml of plasma by the Genomic Mini AX Body Fluids kit (A&A Biotechnology, Gdynia, Poland).

Peripheral blood (12 mL) was collected into K3EDTA tubes (Becton Dickinson, Franklin Lakes, NJ, USA). Plasma was separated within an hour by double centrifugation at 300× g and 1000× g, both at 4 °C for 10 min. DNA was extracted (according to the manufacturer’s instructions) from 1 mL of plasma by the Genomic Mini AX Body Fluids kit (A&A Biotechnology, Gdynia, Poland). Each measurement consisted of a standard curve of three dilutions of plasmid construct containing HPV16 genome, negative control and a samples. For HPV16 detection, reaction was performed using primers and probe set for the HPV16 genome. PCR reactions were performed using the Bio-Rad CFX96 qPCR instrument (Bio-Rad Laboratories, Hemel Hempstead, UK). If HPV16 was found, its presence would be confirmed with a second independent DNA isolation.

Peripheral blood (12 mL) was collected in K3EDTA tubes (Becton–Dickinson, Franklin Lakes, NJ, USA). Plasma was separated within an hour by double centrifugation at 300× g and 1000× g, both at 4 °C for 10 min. DNA was extracted (according to the manufacturer’s instructions) from 1 mL of plasma using the Genomic Mini AX Body Fluids Kit (A&A Biotechnology, Gdynia, Poland). Each measurement consisted of a standard curve of three dilutions of a plasmid construct containing the HPV16 genome, a negative control, and a sample. For HPV16 detection, a reaction was performed using primers and a probe set for the HPV16 genome. PCR reactions were performed using the Bio-Rad CFX96 qPCR instrument (Bio-Rad Laboratories, Hemel Hempstead, UK). If HPV16 was found, its presence was confirmed with a second independent DNA isolation.