FIELD: biotechnology, genetic engineering.
SUBSTANCE: inventions represent sets of primers, pairs of primers and individual primers used in detection and/or identification of transgene DNA sequences in vegetable material and products comprising thereof, methods for detection and/or identification with their using and a device for realization of one of methods. Primers are complementary to 5 typical marker genes and regulatory DNA sequences - gus gene from microorganism E. coli, nptII gene from transposon Tn5, 35S-promoter site in cauliflower mosaic virus, terminator sites in nos gene and ocs gene from A. tumefaciens that comprises most often in genetic constructions of the most abundant in the world market of transgene plant strains and have nucleotide sequences given in SEQ ID Nos. 1-10. Methods involve DNA extraction from vegetable material and products comprising thereof, carrying out asymmetric or symmetric polymerase chain reaction (PCR), among them the multiplex reaction, with taking part of extracted DNA and set of primers, separation of amplified reaction products and their following detection and/or identification by their hybridization with oligonucleotides immobilized on biological microchip or by gel-electrophoresis method, or by using amplifying agent in real time regimen. Original selection of primers and addition of correcting nucleotide exchanges in their sequences provide similarity in their thermodynamic parameters and complete compatibility in sets that provides their effective working in the similar reaction conditions and enhance specificity, sensitivity and reliability of methods for detection and/or identification of transgene DNA sequences with their sharing significantly. Using in method in hybridization analysis of PCR results with specially developed biological microchips BM "Tressigen"-aminated glasses with oligonucleotides immobilized directly on their surface with nucleotide sequences given in SEQ ID Nos. 11-15, and apparatus-program complex "Degmigen" enhances specificity, economy and technological effectiveness of the process significantly. Inventions can be used in monitoring biological safety of nutrient, fodder foodstuffs and other goods of mass consumption.
EFFECT: improved method for analysis, valuable properties of primers.
54 cl, 5 dwg, 1 tbl, 5 ex
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Authors
Dates
2005-12-10—Published
2004-09-15—Filed