Chelating resin-based extraction of DNA from dental pulp and sex determination from incinerated teeth with Y-chromosomal alphoid repeat and short tandem repeats.
A procedure utilizing Chelex 100, chelating resin, was adapted to extract DNA from dental pulp. The procedure was simple and rapid, involved no organic solvents, and did not require multiple tube transfers. The extraction of DNA from dental pulp using this method was as efficient, or more so, than using proteinase K and phenol-chloroform extraction. In this study, the Chelex method was used with amplification and typing at Y-chromosomal loci to determine the effects of temperature on the sex determination of the teeth. The extracted teeth were incinerated in a dental furnace for 2 minutes at 100 degrees C, 200 degrees C, 300 degrees C, 400 degrees C, and 500 degrees C. After the isolation of DNA from the dental pulp by the Chelex method, alphoid repeats, and short tandem repeats, the human Y chromosome (DYZ3), DYS19, SYS389, DYS390, and DYS393 could be amplified and typed in all samples incinerated at up to 300 degrees C for 2 minutes. The DYS389 locus in some samples could not be amplified at 300 degrees C for 2 minutes. An autopsy case is described in which genotypings of DYS19, DYS390, and DYS393 from dental pulp obtained from a burned body were needed. The data presented in this report suggest that Chelex 100-based DNA extraction, amplification, and typing are possible in burned teeth in forensic autopsy cases.
Tsuchimochi
Tsukasa
T
Department of Oral Surgery, Nagoya City University Hospital, Nagoya City University Medical School, Japan.
Iwasa
Mineo
M
Maeno
Yoshitaka
Y
Koyama
Hiroyoshi
H
Inoue
Hiroyuki
H
Isobe
Ichiro
I
Matoba
Ryoji
R
Yokoi
Motoo
M
Nagao
Masataka
M
D002614:Chelating Agents; D041322:Chromosomes, Human, Y / Q000235:genetics*; D004247:DNA / Q000032:analysis / Q000302:isolation & purification*; D016172:DNA Fingerprinting / Q000379:methods; D003782:Dental Pulp / Q000737:chemistry*; D005390:Fires; D005553:Forensic Dentistry / Q000379:methods*; D006801:Humans; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid; D012117:Resins, Synthetic; D012732:Sex Determination Analysis / Q000379:methods
D002614:Chelating Agents; D012117:Resins, Synthetic; C024997:Chelex 100; D004247:DNA