Optimization of multiple PCR reaction system in flue-cured tobacco leaf

Using the single factor and orthogonal experiments with the flue-cured tobacco varieties of zhongyan100 as raw material and two SCAR primers to study the multiple PCR reaction system in the interaction between the main ingredients of flue-cured tobacco leaves,the best suitable multiple PCR reaction system was established for roast tobacco including DNA template 45 ng, Mg²⁺ concentration 2.50 mmol/L, dNTP concentration 0.40 mmol/L and Taq enzyme dosage 4.00 U on the 20 μL system. The stripe of the system is clear and it can shorten the identification time, laying a foundation for using multiple PCR technology to identify flue-cured tobacco varieties.