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Estimating the integrity of aged DNA samples by CE
Author(s) -
Fattorini Paolo,
Marrubini Giorgio,
Ricci Ugo,
Gerin Fabio,
Grignani Pierangela,
Sorçaburu Cigliero Solange,
Xamin Andrea,
Edalucci Elisabetta,
La Marca Giancarlo,
Previderé Carlo
Publication year - 2009
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200900283
Subject(s) - uracil , cytosine , agarose gel electrophoresis , dna , chromatography , chemistry , formaldehyde , agarose , microbiology and biotechnology , biology , biochemistry
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, DN ase I and RN ase 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.