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.

Estimating the integrity of aged DNA samples by CE

MARRUBINI BOULAND, GIORGIO CARLO;GRIGNANI, PIERANGELA;PREVIDERE', CARLO
2009-01-01

Abstract

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.
2009
Sì, ma tipo non specificato
Inglese
Internazionale
STAMPA
30
22
3986
3995
10
Ancient DNA; CE; DNA damage; PCR fidelity; STR typing
10
info:eu-repo/semantics/article
262
Fattorini, P; MARRUBINI BOULAND, GIORGIO CARLO; Ricci, U; Gerin, F; Grignani, Pierangela; Sorçaburu Cigliero, S; Xamin, A; Edalucci, E; La Marca, G; P...espandi
1 Contributo su Rivista::1.1 Articolo in rivista
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11571/208583
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