Agilent 2100 bioanalyzer expert high sensitivity dna assay

A 2 µl aliquot of AMELX-allele and AMELY-allele amplicons generated from positive controls and sex-unknown samples were analyzed by Agilent 2100 Bioanalyzer Expert High Sensitivity DNA Assay (the Bioanalyzer assay) with the High Sensitivity DNA Marker 35/10380 bp, using the Agilent 2100Bioanalyzer (Agilent Technologies Inc.) at the Genomic Services Unit in Langebio, Cinvestav, Mexico.

Bone and teeth samples were processed in a type II B2 biological security hood, located inside the ultraclean room. The outer surface of the bone or teeth was UV-irradiated during 15 minutes and a dental drill was used to remove 0.1 g of powdered material. Powdered samples were added to 5 ml of extraction buffer (0.5 M Tris-HCl pH 8.0–8.5, 0.5 M EDTA pH 8.0, 0.5% SDS and 0.5 mg Proteinase K) and incubated at 37°C for 24 hrs, followed by phenol/chloroform DNA extraction [96] (link). Aliquots of crude DNA extracts were analyzed with an Agilent 2100 Bioanalyzer Expert High Sensitivity DNA Assay to quantify and assess the quality of extracted DNA.
The fresh blood samples (3 ml) were collected in EDTA vacutainer tubes. Total genomic DNA was isolated using a standard non-organic method and diluted to obtain a working concentration of 2.5 ng/µl. All blood samples were processed in a post-PCR laboratory.