| 摘要 |
FIELD: chemistry.SUBSTANCE: invention relates to biotechnology. The method for multiplex detection of Aeromonas and Flavobacterium genus representatives involves performing a polymerase chain reaction in a single step using two pairs of synthetic primers to gene sections of the small subunit of ribosomal RNA (16S rRNA) in real time to the gene section of the small subunit of rRNA Aeromonas: A1 - 5'-TTCGGGCCTTGCGCGATTGG-3' and A2 - 5'-CGTGCTGGCAACAAAGGACAG-3', which facilitate amplification of a 920 bps fragment with melting point of 85.21±0.05°C and to the gene section of the small subunit of rRNA Flavobacterium F1 - 5'-CGATGGATACTAGCTGTTGGG-3' and F2 - 5'-GACGACAACCATGCAGCACC-3', which enable formation of a 242 bps fragment with melting point of 81.23±0.50°C. Thermal cyclers are used according to the program: preliminary denaturation at temperature of 95°C for 5 min, 40 amplification cycles at 95°C - 15 s, 66°C - 60 s; analysis of detection results and determining unknown microorganisms from the presence or absence of specific peaks of the amplified DNA.EFFECT: method provides fast, simple and accurate detection of Aeromonas and Flavobacterium genus representatives in an analysed sample.2 cl, 3 dwg |
