Validation of mitochondrial DNA sequencing for forensic casework analysis

Abstract

Two sets of studies were performed to evaluate the forensic utility of sequencing human mitochondrial DNA (mtDNA) derived from various tissues and amplified by the polymerase chain reaction (PCR). Sequencing was performed on a Perkin-Elmer/Applied Biosystems Division (PE/ABD) automated DNA sequencer (model 373A). The first set of experiments included typical validation studies that had previously been conducted on forensic DNA markers, such as: chemical contaminant effects on DNA from blood and semen and the effect of typing DNA extracted from body fluid samples deposited on various substrates. A second set of experiments was performed strictly on human hair shafts. These studies included typing mtDNA from hairs that were: (1) from different body areas, (2) chemically treated, (3) from deceased individuals, and (4) deliberately contaminated with various body fluids. The data confirm that PCR-based mtDNA typing by direct automated sequencing is a valid and reliable means of forensic identification.

The DNA extraction from the pulp dentine complex of both with and without carious

“…Hypervariable regions (HVRI: L15997-H16395; HVRII: L047-H408) were amplified with primers and amplification conditions according to Wilson et al [5], using Taq Gold. Amplified fragments were purified by MicroSpin Sephadex G50.…”

Mutations in the mitochondrial DNA D-Loop region occur frequently in adenocarcinoma in Barrett's esophagus

“…DNA was extracted from carefully selected specimens as previously described (Schneider et al, 2000). Two sets of published primers L 15997/H 16401 and L 048/H 408 were purchased (MWG Biotech, Ebersberg, Germany) and used to amplify the HV1 and HV2 regions, respectively (Wilson et al, 1995). Thermal cycling conditions were 5 min at 948C, followed by 32 cycles of 948C for 1 min, 608C for 1 min, and 728C for 1 min.…”

Genetic identification of putative remains of the famous astronomer Nicolaus Copernicus

“…(i) PCR amplification was performed using previously described primer pairs (L15997-H16236 and L16159-H16401 (HVI); L48-H285 and L172-H408 (HVII) (24). Amplification was performed in GenAmp 9700 thermocycler (Applied Biosystems) in a total volume of 10 L. The reaction mixture contained 5 L of Qiagen multiplex PCR kit (Qiagen), 1 L of PCR primers, 1 L of Q solution, and 3 L of template DNA.…”