Triplex PCR detection kit for Escherichia coli serotypes O26, O45 and O121 and primer set thereof
A serotype detection, Escherichia coli technology, applied in recombinant DNA technology, DNA/RNA fragment, microbial determination/inspection, etc., can solve problems such as laborious and time-consuming, and achieve the effect of low cost, simple operation and great economic benefits
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[0029] Design of Primer Sets for Detection of Escherichia coli O26, O45, O121 Serotypes
[0030] Three pairs of serotype-specific primers were designed. The 159bp, 241bp and 632bp serotype-specific fragments of O26, O45 and O121 were respectively amplified according to the designed primer set.
[0031] 2. Preparation of Escherichia coli O26, O45, O121 serotype detection kits
[0032] (1) Preparation of E. coli template
[0033] Escherichia coli strains were inoculated into VIB liquid medium and enriched overnight at 37°C. Take 1mL of bacterial liquid each to make a template for later use.
[0034] (2) Preparation of specific primers
[0035]The sizes of target fragments amplified by specific primers for Escherichia coli O26, O45 and O121 detected by this kit are 159bp (O26), 241bp (O45) and 632bp (O121), respectively. The concentration of each primer is 10uM, and the primer dilution is stored at -20°C for future use, avoiding repeated freezing and thawing; it can be artif...
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