[Objective] The study aimed to introduce a rapid and effective method that is suitable for extracting genomic DNA from animal and plant. [ Method ] The genomic DNAs were extracted from tender leaves of 24 peanut cuhiv...[Objective] The study aimed to introduce a rapid and effective method that is suitable for extracting genomic DNA from animal and plant. [ Method ] The genomic DNAs were extracted from tender leaves of 24 peanut cuhivars and from the liver, lung and kidney of white mouse through the specifically modified CTAB method. The DNAs were run on agarose gel, next detected by DNA/Protein analyzer. Finally PCR amplification was conducted to detect the quality of DNAs extracted using the modified CTAB method. [ Result] The clear and orderly bands were observed in gel detection, and the values of OD200/OD200 for DNAs extracted via modified CTAB method were between 1.77 - 1.83. The DNAs performed well in PCR amplification. [ Conclusion] The DNAs extracted by modified CTAB method could satisfy the requirement of PCR amplification.展开更多
ObjectiveThe aim was to seek for a rapid DNA minipreparation method from wheat leaf. MethodThe total DNA of wheat leaf was extracted using CTAB, SDS and boiling water, separately, with some modifications. Integrity an...ObjectiveThe aim was to seek for a rapid DNA minipreparation method from wheat leaf. MethodThe total DNA of wheat leaf was extracted using CTAB, SDS and boiling water, separately, with some modifications. Integrity and purity of nucleic acids were detected through agarose gel electrophoresis, ultraviolet absorption and PCR. ResultThe DNA extracted by the modified CTAB method had high quality and purity, and was not degraded. Two hundreds of DNA samples could be extracted each workday by per capita using this method; and the PCR detection of wheat transgenic plants showed that amplified bands of target gene were clear, without false-positive, and the test results were satisfactory. The DNA purity and concentration extracted by modified SDS method were not as good as that extracted by modified CTAB method, but it also met the DNA requirements of major molecular research. The DNA quantity extracted by modified boiling method was small and there were a lot of impurities in it, PCR detection of this DNA showed no amplified band. ConclusionModified CTAB method is a simple and rapid method for DNA minipreparation from wheat leaf, and was suitable for PCR amplification and other molecular biology researches.展开更多
Leaves of Acer truncatum ＇ Luhong No. 1 ＇ contain large amounts of polysaccharides and polyphenols, which seriously affect extraction yield and quality of total RNA. In order to explore the appropriate total RNA ext...Leaves of Acer truncatum ＇ Luhong No. 1 ＇ contain large amounts of polysaccharides and polyphenols, which seriously affect extraction yield and quality of total RNA. In order to explore the appropriate total RNA extraction method, total RNA was extracted from leaves of A. truncatum ＇ Luhong No. 1 ＇ with three methods, including kit method, Trizol method and modified CTAB method. The results showed that ODE6o/OD2so and OD^o/OD2ao ratios of total RNA extracted from leaves of A. truncatum ＇ Luhong No. 1 ＇ with kit method were higher than 1.8, with a general yield and certain level of DNA contamination ; OD^o/OD~ and OD^o/OD^o ratios of total RNA extracted with Trizol method were about 1.5, with the lowest yield; OD260/OD280 and OD260/OD230 ratios of total RNA extracted with modified CTAB method were about 2.0, with the highest yield and distinct eleetrophoresis patterns. The results demonstrated that total RNA extracted with modified CTAB method exhibited high yield and purity, which could meet the demands of subsequent molecular biology research. Thus, modified CTAB method is the appro- oriate method for extracting total RNA from leaves of A. truncatum ＇ Luhong No. 1 ＇展开更多
为合理评价污染场地风险,以我国云南省某铜堆浸场下游农田土为例,测定土壤中Cu、Pb、Cd和As等4种重金属总量,采用改进BCR(Bureau of European Communities Reference)连续提取法测定4种重金属赋存形态,分析4种重金属间的相关性,并使用...为合理评价污染场地风险,以我国云南省某铜堆浸场下游农田土为例,测定土壤中Cu、Pb、Cd和As等4种重金属总量,采用改进BCR(Bureau of European Communities Reference)连续提取法测定4种重金属赋存形态,分析4种重金属间的相关性,并使用地累积指数法、次生相与原生相比值法(RSP)和风险评价编码法(RAC)对场地土壤重金属污染和潜在风险程度进行评价。结果发现,重金属Cu、Pb、Cd和As平均的含量分别为39.50、46.98、0.16、82.67mg⋅kg^(-1),较云南省重金属背景值,4种重金属超标率分别为0、83.33%、33.33%和100.00%,其中As污染需要特别关注;提取重金属赋存形态结果显示,土样中Cu、Pb、Cd和As均以残渣态为主,占比为83.14%~95.62%,表明4种重金属均以稳定形态存在于土体中;对比3种评价方法,评价结果存在明显差异,地累积指数Igeo结果显示土壤仅As存在污染超标现象,其余3种重金属均无污染,次生相与原生相比值法结果显示,4种重金属对环境均无污染,而风险评价编码法结果显示,Cd对土壤环境存在低风险,其余3种重金属则无风险。对比发现,仅以污染场地土壤重金属总量作为评价指标会一定程度夸大污染风险,应综合重金属总量及赋存形态含量进行分析。虽然重金属元素污染总量较高,但活跃性重金属含量非常低,向环境中扩散的风险也非常低。因此,研究区重金属污染程度较轻,潜在生态风险为低风险。展开更多
文摘[Objective] The study aimed to introduce a rapid and effective method that is suitable for extracting genomic DNA from animal and plant. [ Method ] The genomic DNAs were extracted from tender leaves of 24 peanut cuhivars and from the liver, lung and kidney of white mouse through the specifically modified CTAB method. The DNAs were run on agarose gel, next detected by DNA/Protein analyzer. Finally PCR amplification was conducted to detect the quality of DNAs extracted using the modified CTAB method. [ Result] The clear and orderly bands were observed in gel detection, and the values of OD200/OD200 for DNAs extracted via modified CTAB method were between 1.77 - 1.83. The DNAs performed well in PCR amplification. [ Conclusion] The DNAs extracted by modified CTAB method could satisfy the requirement of PCR amplification.
基金Supported by Major National Transgenic Breeding Project(2011ZX08002-001)the Agricultural Science and Technology Support Program of Jiangsu Province(BE2011306)Agricultural Science and Technology Independent Innovation Fund ofJiangsu Province[CX(12)2026]~~
文摘ObjectiveThe aim was to seek for a rapid DNA minipreparation method from wheat leaf. MethodThe total DNA of wheat leaf was extracted using CTAB, SDS and boiling water, separately, with some modifications. Integrity and purity of nucleic acids were detected through agarose gel electrophoresis, ultraviolet absorption and PCR. ResultThe DNA extracted by the modified CTAB method had high quality and purity, and was not degraded. Two hundreds of DNA samples could be extracted each workday by per capita using this method; and the PCR detection of wheat transgenic plants showed that amplified bands of target gene were clear, without false-positive, and the test results were satisfactory. The DNA purity and concentration extracted by modified SDS method were not as good as that extracted by modified CTAB method, but it also met the DNA requirements of major molecular research. The DNA quantity extracted by modified boiling method was small and there were a lot of impurities in it, PCR detection of this DNA showed no amplified band. ConclusionModified CTAB method is a simple and rapid method for DNA minipreparation from wheat leaf, and was suitable for PCR amplification and other molecular biology researches.
基金Supported by Project of Agricultural Fine Varieties of Shandong Province[LKNZ(2012)No.213]
文摘Leaves of Acer truncatum ＇ Luhong No. 1 ＇ contain large amounts of polysaccharides and polyphenols, which seriously affect extraction yield and quality of total RNA. In order to explore the appropriate total RNA extraction method, total RNA was extracted from leaves of A. truncatum ＇ Luhong No. 1 ＇ with three methods, including kit method, Trizol method and modified CTAB method. The results showed that ODE6o/OD2so and OD^o/OD2ao ratios of total RNA extracted from leaves of A. truncatum ＇ Luhong No. 1 ＇ with kit method were higher than 1.8, with a general yield and certain level of DNA contamination ; OD^o/OD~ and OD^o/OD^o ratios of total RNA extracted with Trizol method were about 1.5, with the lowest yield; OD260/OD280 and OD260/OD230 ratios of total RNA extracted with modified CTAB method were about 2.0, with the highest yield and distinct eleetrophoresis patterns. The results demonstrated that total RNA extracted with modified CTAB method exhibited high yield and purity, which could meet the demands of subsequent molecular biology research. Thus, modified CTAB method is the appro- oriate method for extracting total RNA from leaves of A. truncatum ＇ Luhong No. 1 ＇
文摘为合理评价污染场地风险,以我国云南省某铜堆浸场下游农田土为例,测定土壤中Cu、Pb、Cd和As等4种重金属总量,采用改进BCR(Bureau of European Communities Reference)连续提取法测定4种重金属赋存形态,分析4种重金属间的相关性,并使用地累积指数法、次生相与原生相比值法(RSP)和风险评价编码法(RAC)对场地土壤重金属污染和潜在风险程度进行评价。结果发现,重金属Cu、Pb、Cd和As平均的含量分别为39.50、46.98、0.16、82.67mg⋅kg^(-1),较云南省重金属背景值,4种重金属超标率分别为0、83.33%、33.33%和100.00%,其中As污染需要特别关注;提取重金属赋存形态结果显示,土样中Cu、Pb、Cd和As均以残渣态为主,占比为83.14%~95.62%,表明4种重金属均以稳定形态存在于土体中;对比3种评价方法,评价结果存在明显差异,地累积指数Igeo结果显示土壤仅As存在污染超标现象,其余3种重金属均无污染,次生相与原生相比值法结果显示,4种重金属对环境均无污染,而风险评价编码法结果显示,Cd对土壤环境存在低风险,其余3种重金属则无风险。对比发现,仅以污染场地土壤重金属总量作为评价指标会一定程度夸大污染风险,应综合重金属总量及赋存形态含量进行分析。虽然重金属元素污染总量较高,但活跃性重金属含量非常低,向环境中扩散的风险也非常低。因此,研究区重金属污染程度较轻,潜在生态风险为低风险。
