[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
Along with the emergence and ma- turity of 100Gbit/s technologies, numbers of trial and commercial 100Gbit/s Wavelength Division Multiplexing （WDM） systems have been established worldwide. The research- oriented lab...Along with the emergence and ma- turity of 100Gbit/s technologies, numbers of trial and commercial 100Gbit/s Wavelength Division Multiplexing （WDM） systems have been established worldwide. The research- oriented lab tests of 100Gbit/s WDM systems were done by China Unicorn in 2012. The 100Gbit/s technologies of polarization multi- plex-quadrature phase shift keying with co- herent detection from multiple vendors have been studied. There are some further issues to be specified before the commercial application of 100Gbit/s WDM system is implemented in large scale in backbone network, including optical pertbnnance parameters, optical per- lbrmance monitoring of 100Gbit/s system, protection mechanism, selection of client in- terfhce and comprehensive Operation, Ad- ministration and Maintenance （OAM）.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supported by the China Unicom under the project of the research on 100Gbit/s high bitrate transport technology
文摘Along with the emergence and ma- turity of 100Gbit/s technologies, numbers of trial and commercial 100Gbit/s Wavelength Division Multiplexing （WDM） systems have been established worldwide. The research- oriented lab tests of 100Gbit/s WDM systems were done by China Unicorn in 2012. The 100Gbit/s technologies of polarization multi- plex-quadrature phase shift keying with co- herent detection from multiple vendors have been studied. There are some further issues to be specified before the commercial application of 100Gbit/s WDM system is implemented in large scale in backbone network, including optical pertbnnance parameters, optical per- lbrmance monitoring of 100Gbit/s system, protection mechanism, selection of client in- terfhce and comprehensive Operation, Ad- ministration and Maintenance （OAM）.
