短串联重复序列(short tandem repeat,STR)是存在于人类基因组中的一类具有长度多态性的DNA序列,由含2~6个碱基对的重复单位串联构成。DNA STR分型检验是当前法庭科学进行个体识别和亲缘鉴定的主要依据。STR分型标准物质是DNA STR检验...短串联重复序列(short tandem repeat,STR)是存在于人类基因组中的一类具有长度多态性的DNA序列,由含2~6个碱基对的重复单位串联构成。DNA STR分型检验是当前法庭科学进行个体识别和亲缘鉴定的主要依据。STR分型标准物质是DNA STR检验量值溯源的基础和关键物质,对其进行研究将极大推动我国法庭科学DNA STR检验的标准化进程。以STR基因座D6S1043为例,介绍人类基因组中STR序列的质粒DNA标准物质的制备过程。首先,合成包含13个重复单元([ATCT]13)的D6S1043序列(定义为D6S1043-1)并将其与pUC57载体连接构建重组质粒,制备质粒溶液。其次,通过酶切鉴定、Sanger测序、多种STR分型试剂盒的分型鉴定等方法检验DNA序列的准确性。再次,采用微滴式数字PCR方法(droplet digital PCR,ddPCR)检测质粒浓度,评估质粒溶液的均匀性和稳定性。最后,由8家实验室协作,测定浓度标准值,综合分析均匀性检验、稳定性检验、定值过程等引入的不确定度,得到总不确定度;由6家实验室协作,测定D6S1043-1的分型值。结果表明:该标准物质的均匀性、稳定性良好,在-20℃可保存6个月,在4℃可保存14 d;核酸拷贝数为(7.7±1.2)×10^3 copies·μL^-1;在D6S1043基因座上的分型值为13。人D6S1043-1型STR质粒DNA标准物质[编号:GBW(E)091072]是我国法庭科学DNA检验领域首批有证标准物质之一,其研制过程为其他STR基因座的质粒DNA标准物质的研制提供了一定的借鉴。展开更多
An experimental setup of laser-induced graphite plasma was built and the spectral characteristics and properties of graphite plasma were studied. From the temporal behavior of graphite plasma, the duration of CN parti...An experimental setup of laser-induced graphite plasma was built and the spectral characteristics and properties of graphite plasma were studied. From the temporal behavior of graphite plasma, the duration of CN partials （B2 ∑＋ →-X2 ∑＋） emission was two times longer than that of atomic carbon, and all intensities reached the maximum during the early stage from 0.2 μs to 0.8 μs. The electron temperature decreased from 11807 K to 8755 K, the vibration temperature decreased from 8973 K to 6472 K, and the rotational temperature decreased from 7288 K to 4491 K with the delay time, respectively. The effect of the laser energy was also studied, and it was found that the thresholds and spectral characteristics of CN molecular and C atomic spectroscopy presented great differences. At lower laser energies, the electron excited temperature, the electron density, the vibrational temperature and rotational temperature of CN partials increased rapidly. At higher laser energies, the increasing of electron excited temperature and electron density slow down, and the vibrational temperature and rotational temperature even trend to saturation due to plasma shielding and dissociation of CN molecules. The relationship among the three kinds of temperatures was Telec〉Tvib〉Trot at the same time. The electron density of the graphite plasma was in the order of 1017 cm-3 and 1018 cm-3.展开更多
文摘短串联重复序列(short tandem repeat,STR)是存在于人类基因组中的一类具有长度多态性的DNA序列,由含2~6个碱基对的重复单位串联构成。DNA STR分型检验是当前法庭科学进行个体识别和亲缘鉴定的主要依据。STR分型标准物质是DNA STR检验量值溯源的基础和关键物质,对其进行研究将极大推动我国法庭科学DNA STR检验的标准化进程。以STR基因座D6S1043为例,介绍人类基因组中STR序列的质粒DNA标准物质的制备过程。首先,合成包含13个重复单元([ATCT]13)的D6S1043序列(定义为D6S1043-1)并将其与pUC57载体连接构建重组质粒,制备质粒溶液。其次,通过酶切鉴定、Sanger测序、多种STR分型试剂盒的分型鉴定等方法检验DNA序列的准确性。再次,采用微滴式数字PCR方法(droplet digital PCR,ddPCR)检测质粒浓度,评估质粒溶液的均匀性和稳定性。最后,由8家实验室协作,测定浓度标准值,综合分析均匀性检验、稳定性检验、定值过程等引入的不确定度,得到总不确定度;由6家实验室协作,测定D6S1043-1的分型值。结果表明:该标准物质的均匀性、稳定性良好,在-20℃可保存6个月,在4℃可保存14 d;核酸拷贝数为(7.7±1.2)×10^3 copies·μL^-1;在D6S1043基因座上的分型值为13。人D6S1043-1型STR质粒DNA标准物质[编号:GBW(E)091072]是我国法庭科学DNA检验领域首批有证标准物质之一,其研制过程为其他STR基因座的质粒DNA标准物质的研制提供了一定的借鉴。
基金supported by National Natural Science Foundation of China(No.61205149)Scientific Research Foundation for the Returned Overseas Chinese Scholars of State Education Ministry,Science Research Funds of Chongqing Municipal Education Commission(KJ1500436)+2 种基金Scientific and Technological Talents Training Project of Chongqing(CSTC2013kjrc-qnrc40002)Key Project of Foundation and Advanced Technology Research Project of Chongqing(CSTC2015jcyj B0358)Visiting Scholarship of State Key Laboratory of Power Transmission Equipment & System Security and New Technology(2007DA10512714409)
文摘An experimental setup of laser-induced graphite plasma was built and the spectral characteristics and properties of graphite plasma were studied. From the temporal behavior of graphite plasma, the duration of CN partials （B2 ∑＋ →-X2 ∑＋） emission was two times longer than that of atomic carbon, and all intensities reached the maximum during the early stage from 0.2 μs to 0.8 μs. The electron temperature decreased from 11807 K to 8755 K, the vibration temperature decreased from 8973 K to 6472 K, and the rotational temperature decreased from 7288 K to 4491 K with the delay time, respectively. The effect of the laser energy was also studied, and it was found that the thresholds and spectral characteristics of CN molecular and C atomic spectroscopy presented great differences. At lower laser energies, the electron excited temperature, the electron density, the vibrational temperature and rotational temperature of CN partials increased rapidly. At higher laser energies, the increasing of electron excited temperature and electron density slow down, and the vibrational temperature and rotational temperature even trend to saturation due to plasma shielding and dissociation of CN molecules. The relationship among the three kinds of temperatures was Telec〉Tvib〉Trot at the same time. The electron density of the graphite plasma was in the order of 1017 cm-3 and 1018 cm-3.
