Sample degradation leads to false-positive copy number variation calls in multiplex real-time polymerase chain reaction assays.
Anal Biochem
; 386(2): 288-90, 2009 Mar 15.
Article
em En
| MEDLINE
| ID: mdl-19121619
ABSTRACT
The recent implication of genomic copy number variations (CNVs) in multiple human genetic disorders has led to increased interest in CNV discovery technologies. There is a growing consensus that, in addition to the method used for detection, at least one additional technology should be employed for validation. Real-time quantitative polymerase chain reaction (qPCR) analysis, incorporating a normal (2N) copy number standard, is commonly used as a means of validating CNVs. Whereas it has previously been reported that formalin-fixed paraffin-embedded (FFPE) DNA samples can yield spurious CNV calls in real-time qPCR assays, here we report that sample degradation under standard laboratory storage conditions generates a significant increase in false-positive CNV results. Results suggest the possibility of biased degradation among genomic regions and emphasize the need to assess sample integrity immediately prior to real-time qPCR experiments.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Variação Genética
/
Genoma Humano
/
Reação em Cadeia da Polimerase
Tipo de estudo:
Diagnostic_studies
Limite:
Humans
Idioma:
En
Ano de publicação:
2009
Tipo de documento:
Article