[Microarray analytic system for multiplex analysis by real-time polymerase chain reaction with reagents immobilized in microreactors].
A microarray analytic system that uses a silicon chip with immobilized in microreactor test-system for multiplex analysis of DNA by real-time polymerase chain reaction (RT-PCR) was developed and optimized. We suggested the method of immobilization of PCR-components of a test-system, chose the stabilizer, and conducted the optimization of the composition of reaction mixture to achieve permanent stability of a microarray. We conducted optimization of preparation of samples using magnetic sorbent and indicated that, with 2.6 x 10(4) copies/ml, 60 min are necessary to obtain positive identification including time for preparation of model probes. The abilities of the created system were demonstrated on the example of microarray analysis of samples with different content of DNA, low absolute limits of identification (20 DNA copies in microreactor), and high reproducibility of the analysis.
D V
DV
Navolotskiĭ
A V
AV
Perchik
I A
IA
Mark'ianov
A A
AA
Ganeev
M N
MN
Sliadnev
D000327:Adsorption; D018744:DNA, Plant / Q000032:analysis*; D005612:Freeze Drying; D018628:Gene Dosage; D007202:Indicators and Reagents / Q000737:chemistry; D056656:Lab-On-A-Chip Devices; D059346:Magnets; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results; D012639:Seeds / Q000737:chemistry; D012825:Silicon / Q000737:chemistry; D003313:Zea mays / Q000737:chemistry*
D018744:DNA, Plant; D007202:Indicators and Reagents; D012825:Silicon