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《Grassland and Turf》 2009-01
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Establishment and optimization of ISSR-PCR reaction system of centipedegrass

YANG Shui-lian,LIU Wei-dong,MA Tao,SHU Bi-chao,LIU Rui-feng,XIE Peng(Central South University of Forestry and Technology,Changsha 410004,China)  
Centipedegrass was used to study the affecting factors of ISSR-PCR amplification.A suitable reaction system was established under the following conditions: 20 ng template DNA,0.4 μmol/L random primers,0.5 mmol/L Mg2+,0.05 mmol/L dNTPs,0.2 U TaqDNA polymerase;and 2.0 μL 10×buffer in 20 μL total reaction solution;and the optimized annealing temperature was 55 ℃.The PCR procedure was as follows: pre-denaturizing at 94 ℃ for 5 min,denaturizing at 94 ℃ for 45 s,annealing at 55 ℃ for 45 s,extension at 72 ℃ for 1.5 min,reaction of 35 cycles and re-extension at 72 ℃ for 7 min.
【Fund】: 湖南省科技厅项目(20-07NK3118);; 国家林业局“亚热带森林植物种质资源保存库建设”资助
【CateGory Index】: S688.4
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