Optimization of SRAP-PCR System for Tomato Based on Orthogonal Design
GUO Cai-jie1, 2, HOU Li-xia1*, CUI Na2*, HAN Ming-li2
1Institute of Vegetables, Shandong Academy of Agricultural Sciences, Key Lab for Biology of Greenhouse Vegetable of Shandong Province, National Center for Vegetable Improvement（Shandong）, Jinan 250100, Shandong, China; 2Biological Science and Technology College, Shenyang Agricultural University, Shenyang 110161, Liaoning, China
Abstract：The orthogonal design was used to optimize SRAP-PCR amplification system at 4 levels of 5 factors in tomato（Lycopersion esculentum Mill.）（DNA template, primer, Mg2+, dNTPs and Taq DNA polymerase）. The results showed that the order of each factor in different levels affected the result of PCR was: primer＞Taq DNA polymerase＞dNTPs＞DNA template＞Mg2+. The most suitable SRAP-PCR reaction system for tomato was total 20 μL containing 15 ng DNA template, 0.75 μmol?L-1 primer, 2.0 mmol?L-1 Mg2+, 0.125 mmol?L-1 dNTPs and 1.0 U Taq DNA polymerase.
