After the extraction, each dental piece was conserved in sterile
gauze at 4 8C until its subsequent processing. A mechanical manual
and fast fragmentation in one hit was applied to each piece,
collecting the pulp tissue from the pulp chamber and the root canal
without pollutants (Fig. 1). Pulp tissue was collecting in an
eppendorf tube with 1 ml of saline and conserved refrigerated until
DNA extraction.
DNA from the total pulp material was extracted by the
commercial QIAamp1 Mini Kit (Qiagen, Hilden, Germany)
following the manufacturer instructions for tissues. Human
DNA concentration was determined by the QuantifilerTM System
Human DNA Quantification Kit (Life Technologies, Foster City, CA,
USA), whereas NanoDropTM Lite Spectrophotometer (Thermo-
Scientific) was used to determine the total concentration and the
purity of the samples through the A260/280 coefficient.