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PCR Cloning and Subcloning> | … maculans and L. biglobosa were synthesized and cloned into plasmid pUC57, respectively (Genscript, Jiangsu, China). The resultant recombinant plasmid, termed pUC57-Lm-ITS, and pUC57-Lb-ITS was replicated in E.coli DH5α cells, extracted and purified using the Endotoxin … | Get A Quote |
The fungus Leptosphaeria maculans leading to Phoma stem canker (blackleg) of Brassica napus (oilseed rape, canola) produces the phytotoxin sirodesmin PL, which is responsible for major yield losses of oilseed rape worldwide. Polymerase chain reaction (PCR) remains the gold standard diagnostic tool for L. maculans, but the required expensive equipment and long time make it inappropriate for fast field test. Herein, a portable system for rapid assaying L. maculans and L. biglobosa is designed around recombinase polymerase amplification (RPA) with fluorescent probe as the signal indicator, which allowed the real-time assay of amplification performed on a portable device between 37 and 42 °C. The time ... More