Estimating the integrity of aged DNA samples by CE.
A CE/UV method was developed to separate by a micellar system the four DNA bases and other five purinic-pyrimidinic compounds (5-methyl-cytosine, uracil, xanthyne, hypoxanthyne and 5-bromo-uracil). Selectivity, precision, accuracy and sensitivity were assessed and proved to be suitable for the analysis of the primary structure of DNA. This method was adopted to study 16 aged samples including two Egyptian mummies, formaldehyde-fixed paraffin-embedded tissues and other forensic specimens. Lower relative values of the four canonical unmodified DNA bases (uDNAb) and more complex pherograms were found in the aged samples when compared with the modern controls. The results of the CE analysis, together with those obtained by classical molecular methods (agarose gel electrophoresis, DNase I and RNase A assays, and UV spectrophotometry), were finally evaluated for assessing the reliability of STR typing. Since samples with low uDNAb showed no amplification or unreliable STR profiles, the uDNAb value is discussed as a further quality criterion in the evaluation of the genetic data obtained from aged samples.
Fattorini
Paolo
P
Department of Sciences of Public Health, University of Trieste, Italy. fattorin@univ.trieste.it
Marrubini
Giorgio
G
Ricci
Ugo
U
Gerin
Fabio
F
Grignani
Pierangela
P
Cigliero
Solange Sorçaburu
SS
Xamin
Andrea
A
Edalucci
Elisabetta
E
La Marca
Giancarlo
G
Previderé
Carlo
C
D004247:DNA / Q000032:analysis / Q000737:chemistry*; D053836:DNA Degradation, Necrotic; D019075:Electrophoresis, Capillary / Q000379:methods*; D005404:Fixatives; D018732:Forensic Anthropology / Q000379:methods; D005554:Forensic Medicine / Q000379:methods; D005557:Formaldehyde; D006801:Humans; D009106:Mummies; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*; D013056:Spectrophotometry, Ultraviolet; D020080:Tandem Repeat Sequences
D005404:Fixatives; D005557:Formaldehyde; D004247:DNA