Fidelity of polymerase chain reaction-direct sequencing analysis of damaged forensic samples.
作者： Paolo Fattorini , Riccardo Ciofuli , Federica Cossutta , Piero Giulianini , Paolo Edomi
DOI: 10.1002/(SICI)1522-2683(19991101)20:17<3349::AID-ELPS3349>3.0.CO;2-7
关键词:
摘要: Polymerase chain reaction (PCR) direct sequence analysis was performed on aged forensic samples, six or thirteen years old. This method allowed unambiguous genetic typing, but PCR products from such samples showed several artifacts. Control generated ambiguities at a frequency of 1 in 567 bases, the had an error about 30-fold higher. In order to study molecular composition these DNA reversed-phase high performance liquid chromatography (HPLC) performed. Reduced amounts four bases were observed and anomalous peaks found. These analyzed by ionization mass spectrometry identified as oxidation. The sequencing artifacts found be proportional decay templates. Although fidelity is relevant concern damaged our data indicate that risk mistyping circumventable both strands performing replicate amplifications same template.
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