Significance of methods for purification of oligodeoxyribonucleotide probes for the efficiency of gene diagnosis by real-time PCR.
Analysis of the use of real-time PCR with fluorescent registration of results for gene diagnosis of infectious diseases showed that the sensitivity and reliability of quantitative evaluation of DNA targets directly depended on the method of purification of oligonucleotide probes. Chromatographic behavior of synthetic probes carrying various fluorophores and fluorescence quenchers was analyzed. Approaches to optimization of purification methods are proposed enabling elimination of previously undetectable admixtures. The importance of these studies is explained by the need in extending the armory of methods for the development and production of diagnosticums for detection of infectious and hereditary diseases, identification of genetically modified organisms, and for a wide spectrum of research in molecular biology and medicine.
Tatarinova
O N
ON
Institute of Physicochemical Medicine, Russian Ministry of Health.
Lukyanova
T N
TN
Zaitseva
M A
MA
Veremeev
K Y
KY
Karpov
V A
VA
Chuvilin
A N
AN
Petrunin
D D
DD
Pozmogova
G E
GE
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004247:DNA / Q000032:analysis; D006801:Humans; D015345:Oligonucleotide Probes / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*
D015345:Oligonucleotide Probes; D004247:DNA