Orthogonal design

Optimization of SCoT-PCR reaction system in Dactylis glomerata by orthogonal design

B. Zeng, Huang, X., Huang, L. K., Zhang, J., Yan, H. D., Luo, D., Liang, H., and Yuan, Y., Optimization of SCoT-PCR reaction system in Dactylis glomerata by orthogonal design, vol. 14, pp. 3052-3061, 2015.

The effects of 5 factors (template DNA, Mg2+, dNTPs, Taq DNA polymerase, and primer) on the polymerase chain reaction (PCR) were investigated to optimize the start codon targeted polymor­phism (SCoT)-PCR system of Dactylis glomerata L., using an orthogo­nal design L16 (45). A suitable SCoT-PCR system for D. glomerata was established; the 20 μL reaction volume contained 3.0 mM Mg2+, 0.2 mM dNTPs, 1.0 U Taq DNA polymerase, 0.2 μM primer, 20 ng tem­plate DNA, and 2 μL 10X buffer.

Extraction of total DNA and optimization of the RAPD reaction system in Dioscorea opposita Thunb.

G. Q. Wen, Li, J., Liu, X. H., Zhang, Y. S., and Wen, S. S., Extraction of total DNA and optimization of the RAPD reaction system in Dioscorea opposita Thunb., vol. 13, pp. 1339-1347, 2014.

Dioscorea opposita Thunb. has been used as health food and herbal medicinal ingredients in traditional Chinese medicine. In this study, the total DNA of D. opposita Thunb. was extracted using an improved cetyltrimethylammonium bromide (CTAB) method, and the extracted DNA was further used for random amplified polymorphic DNA (RAPD) reaction system by design of the L16 (44) orthogonal diagram.

Optimization of factors affecting Agrobacterium-mediated transformation of Micro-Tom tomatoes

M. Guo, Zhang, Y. L., Meng, Z. J., and Jiang, J., Optimization of factors affecting Agrobacterium-mediated transformation of Micro-Tom tomatoes, vol. 11, pp. 661-671, 2012.

Micro-Tom is the smallest known variety of tomatoes. An orthogonal experimental design L16 (45) was used to optimize Agrobacterium-mediated transformation of cotyledon explants of Lycopersicon esculentum cv. Micro-Tom. Four parameters were investigated to determine their effect on transformation frequency: the concentration of bacterial suspension, time of dip in bacterial suspension, co-cultivation time, and concentration of carbenicillin.

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