DNA isolation with manual systems

For manual DNA isolation, two commercially available kits were used: the PureLink Genomic DNA Mini Kit from Invitrogen Carlsbad, CA 92008, United States and the High Pure FFPE DNA Isolation Kit from Roche GmbH, Mannheim, Germany. For each of the 46 included cases, two manual isolations per case were performed according to the manufacturers’ protocols (Table ​(Table11).

DNA isolation protocols

Invitrogen kits were tested at the Pathology Department of Mures County Hospital, while Roche kits were tested at the Molecular Pathology Laboratory of the National Institute of Oncology, Budapest. The same team performed all isolations.

The manual methods (for both kits) were performed on an anion-exchange resin. It is a macroporous silica-based resin with a high density of diethylaminoethyl (DEAE) groups. The purification is based on the interaction between the negatively charged phosphates of the nucleic acid backbone and the positively charged DEAE groups on the surface of the resin[7].