[Objective] This study aimed to establish a multiplex PCR system for de- tecting transgenic ingredients from Citrus. [Method] Based on the pBI121 plasmid sequences published in GenBank and actin gene sequence of Citru...[Objective] This study aimed to establish a multiplex PCR system for de- tecting transgenic ingredients from Citrus. [Method] Based on the pBI121 plasmid sequences published in GenBank and actin gene sequence of Citrus, the primers specific to CaMV35S promoter, NOS promoter, NOS terminator and actin gene were designed, to establish a multiple PCR system which could detect four types of sequences. In addition, orthogonal tests were performed to determine the optimal concentrations of all the components in PCR reaction system, as well as the optimal PCR cycle parameters. [Result] The optimal PCR reaction system should contain 2.5μl of 10xPCR buffer, 2.0μl of MgCI2 （25 mmol/L）, 2.0 μl of dNTP mixture （2.5 mmol/L of each dNTP）, 1.0 μl of actin gene primers （10μmol/L）, 1.0μl of 35S promoter primers （10 μmol/L）, 1.5 μl of NOS promoter primers （10 μmol/L） and 0.5 μl of NOS terminator primers （10μmol/L）, 0.1 μg of template DNA, 1.25 U of Taq DNA polymerase; ddH20 was added to the total reaction system of 25μl. The PCR reaction program consisted of pre-denaturing at 94℃ for 5 min; 31 cycles of denaturing at 94℃ for 30 s, annealing at 64.1℃ for 45 s and extension at 72℃ for 50 s; final extension at 72℃ for 10 min. The reaction system optimized with the orthogonal tests could detect as less as 0.1% transgenic component in the tested samples. [Conclusion] The MPCR detection system established in this study can meet the requirements in theory for detecting the genetically modified ingredients in Citrus or the deep-processed products.展开更多
Multi-stand roll forming is a process that has very complicated deformation behaviour and shows significant nonlinearity.In this paper, the sensitivity analysis of parameters for multi-stand roll forming was performed...Multi-stand roll forming is a process that has very complicated deformation behaviour and shows significant nonlinearity.In this paper, the sensitivity analysis of parameters for multi-stand roll forming was performed via a new booting finite element method(FEM) model.Compared with the most of simulation, the new model is more consistent with production process and can account for the effects of roll rotating speed.Based on the model, the process of an open section channel formed with 10 passes was simulated and the sensitivity analysis was conducted with orthogonal experiment design combined FEM model.The multi-stand roll forming process can be efficiently analyzed by the new booting model.And sensitivity analysis shows the hardening exponent plays an important role in controlling the quality of the products.展开更多
基金Supported by the Special Fund for Key Laboratories of Chongqing (CSTC)National Technology Research and Development Program of Ministry of Science and Technology for Countryside Field (863 Program,2011AA100205)+1 种基金Special Fund for Agro-scientific Research in the Public Interest of Ministry of Agriculture of China(201003067)Key Science and Technology Research Program of Ministry of Education of China (109131)~~
文摘[Objective] This study aimed to establish a multiplex PCR system for de- tecting transgenic ingredients from Citrus. [Method] Based on the pBI121 plasmid sequences published in GenBank and actin gene sequence of Citrus, the primers specific to CaMV35S promoter, NOS promoter, NOS terminator and actin gene were designed, to establish a multiple PCR system which could detect four types of sequences. In addition, orthogonal tests were performed to determine the optimal concentrations of all the components in PCR reaction system, as well as the optimal PCR cycle parameters. [Result] The optimal PCR reaction system should contain 2.5μl of 10xPCR buffer, 2.0μl of MgCI2 （25 mmol/L）, 2.0 μl of dNTP mixture （2.5 mmol/L of each dNTP）, 1.0 μl of actin gene primers （10μmol/L）, 1.0μl of 35S promoter primers （10 μmol/L）, 1.5 μl of NOS promoter primers （10 μmol/L） and 0.5 μl of NOS terminator primers （10μmol/L）, 0.1 μg of template DNA, 1.25 U of Taq DNA polymerase; ddH20 was added to the total reaction system of 25μl. The PCR reaction program consisted of pre-denaturing at 94℃ for 5 min; 31 cycles of denaturing at 94℃ for 30 s, annealing at 64.1℃ for 45 s and extension at 72℃ for 50 s; final extension at 72℃ for 10 min. The reaction system optimized with the orthogonal tests could detect as less as 0.1% transgenic component in the tested samples. [Conclusion] The MPCR detection system established in this study can meet the requirements in theory for detecting the genetically modified ingredients in Citrus or the deep-processed products.
基金the National Natural Science Foundation of China (No. 50605043)
文摘Multi-stand roll forming is a process that has very complicated deformation behaviour and shows significant nonlinearity.In this paper, the sensitivity analysis of parameters for multi-stand roll forming was performed via a new booting finite element method(FEM) model.Compared with the most of simulation, the new model is more consistent with production process and can account for the effects of roll rotating speed.Based on the model, the process of an open section channel formed with 10 passes was simulated and the sensitivity analysis was conducted with orthogonal experiment design combined FEM model.The multi-stand roll forming process can be efficiently analyzed by the new booting model.And sensitivity analysis shows the hardening exponent plays an important role in controlling the quality of the products.
