[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
[Objective] This study was to find out a quick,simple,and low-cost method for the extraction of sorghum genomic DNA.[Method] Four plant genomic DNA extraction methods based on CTAB,including liquid nitrogen grinding m...[Objective] This study was to find out a quick,simple,and low-cost method for the extraction of sorghum genomic DNA.[Method] Four plant genomic DNA extraction methods based on CTAB,including liquid nitrogen grinding method（method I）,buffer grinding method（method II）,drying grinding method（method III）and directly grinding method（method IV）,were used to extract the sorghum genomic DNA from leaves;further the quantity and quality of the yielded DNA were detected by gel electrophoresis,SSR-PCR and SRAP-PCR.[Result] These four methods performed no remarkable difference in DNA product.The method I and method II produced DNA with higher purity and better integrity,which,especially from method I,is effective for SRAP-PCR and SSR-PCR.While the DNA extracted via method III and method IV had less integrality and lower purity,and only effective in SSR-PCR.[Conclusion] Enough amount of sorghum genomic DNA to perform tens of PCR could be quickly extracted using all these four methods.The DNA obtained via method I and method II had a broader application spectrum（SRAP,RAPD,ISSR and SSR）than that via method III and method IV which is only proper for PCR targeting small DNA fragments（SSR）.展开更多
[Objective] This study aimed at comparing the four extraction methods of genomic DNA from Clematis fasciculiflora Franch and determining the optimal extraction method for extracting the genomic DNA from Clematis fasci...[Objective] This study aimed at comparing the four extraction methods of genomic DNA from Clematis fasciculiflora Franch and determining the optimal extraction method for extracting the genomic DNA from Clematis fasciculiflora Franch.[Method] Leavies of Clematis fasciculiflora Franch were used as materials for comparing the purity and concentration of extracted DNA and extracting time among the four extraction methods of genomic DNA including improved CTAB method Ⅰ,improved CTAB method Ⅱ,improved CTAB method Ⅲ and improved SDS method.[Result] The four extraction methods could all be successfully used for extracting the genomic DNA from Clematis fasciculiflora Franch.The purity of genomic DNA was the highest using improved CTAB method Ⅰ,with the longest extracting time;while the concentration of genomic DNA was the maximum using the improved SDS method,with the shortest extracting time and relatively low purity;the extracting time of improved CTAB method Ⅲ was the shortest.[Conclusion] This study had established the optimal extraction method for extracting the genomic DNA from Clematis fasciculiflora Franch and supported for the further research using molecular biological methods.展开更多
[ Objective] The aim was to study method for extracting DNA from pteridophyta and provide basis for further study on genetic diversity and taxonomy. [ Method] By changing the dosage of reagent and operating method, CT...[ Objective] The aim was to study method for extracting DNA from pteridophyta and provide basis for further study on genetic diversity and taxonomy. [ Method] By changing the dosage of reagent and operating method, CTAB method for extracting DNA was improved. [ Result] The results showed that the improved CTAB method could extract high-quality DNA from pteridophyta. [ Conclusion] The study improved method for extracting DNA from pteridophyta.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
[Objective] The research aimed to isolate flanking sequences adjacent to the transgenic T-DNA in Brassica napus by an improved inverse PCR method.[Method] Using single clone of transgenic FS4 in Brassica napus as the ...[Objective] The research aimed to isolate flanking sequences adjacent to the transgenic T-DNA in Brassica napus by an improved inverse PCR method.[Method] Using single clone of transgenic FS4 in Brassica napus as the research materials,total DNA was extracted from transgenic Brassica napus by using modified CTAB method.After enzyme digestion and purification,self-joining was made.Two circles of nested PCR and the sequence alignment were carried out.[Result] A fragement with the size of 4.0 kb was amplified ...展开更多
[Objective] The present study aimed to establish a rapid method for the isolation of small amount DNA from citrus.[Method] By using the improved CTBA method,the genomic DNA was extracted respectively from 20,10,5 and ...[Objective] The present study aimed to establish a rapid method for the isolation of small amount DNA from citrus.[Method] By using the improved CTBA method,the genomic DNA was extracted respectively from 20,10,5 and 2.5 mg hybrid embryos of citrus,and then the DNA quality was detected and followed by SSR verification.[Result] The method was very simple and rapid,which needed less materials.In addition,the isolated DNA showed good purity with the OD260/OD280 of 1.8-2.1,and could meet the requirement for PCR-based technology,such as SSR,etc..[Conclusion] The method could be used for rapid extraction of small amount of genomic DNA from citrus.展开更多
An effective method for isolation of total RNA from pulps of mainly deciduous fruit trees was discussed in this paper. Here we shown that total RNA was successfully isolated from pulps of apple, peach and strawberry, ...An effective method for isolation of total RNA from pulps of mainly deciduous fruit trees was discussed in this paper. Here we shown that total RNA was successfully isolated from pulps of apple, peach and strawberry, but not grape berry via the effective CTAB method modified by ZENG, et al. However, total RNA was isolated from grape berry according to ZENG＇s protocols with a modification including 16% of β-Mercatoethanol and 3% of PVP in extraction buffer, 1/3 （V/V） of 5mol/L potassium acetate （KAC PH4.8） in supernatant. The detection of the isolated total RNA above using ultraviolet spectrophoto-metry, electrophoresis, reverse transcription and RT-PCR shown that the CTAB and its modifications is a preferred method for isolation of high-quality total RNA from fruit pulps.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 Key Technology R&D Program of Tianjin(10ZCKFNC00100)National Key Technology R&D Program(2007BAD42B03)~~
文摘[Objective] This study was to find out a quick,simple,and low-cost method for the extraction of sorghum genomic DNA.[Method] Four plant genomic DNA extraction methods based on CTAB,including liquid nitrogen grinding method（method I）,buffer grinding method（method II）,drying grinding method（method III）and directly grinding method（method IV）,were used to extract the sorghum genomic DNA from leaves;further the quantity and quality of the yielded DNA were detected by gel electrophoresis,SSR-PCR and SRAP-PCR.[Result] These four methods performed no remarkable difference in DNA product.The method I and method II produced DNA with higher purity and better integrity,which,especially from method I,is effective for SRAP-PCR and SSR-PCR.While the DNA extracted via method III and method IV had less integrality and lower purity,and only effective in SSR-PCR.[Conclusion] Enough amount of sorghum genomic DNA to perform tens of PCR could be quickly extracted using all these four methods.The DNA obtained via method I and method II had a broader application spectrum（SRAP,RAPD,ISSR and SSR）than that via method III and method IV which is only proper for PCR targeting small DNA fragments（SSR）.
基金Supported by Applied Basic Research Project of Yunnan Province(2010ZC089)the948Project of National Forestry Bureau(2008-4-11)+1 种基金Sharing Platform Project of Provincial and Ministerial Key Subject,Key Laboratory and School Laboratory of Provincial Colleges and Universities in Yunnan ProvinceScience and Technology Innovation Fund of Southwest Forestry University~~
文摘[Objective] This study aimed at comparing the four extraction methods of genomic DNA from Clematis fasciculiflora Franch and determining the optimal extraction method for extracting the genomic DNA from Clematis fasciculiflora Franch.[Method] Leavies of Clematis fasciculiflora Franch were used as materials for comparing the purity and concentration of extracted DNA and extracting time among the four extraction methods of genomic DNA including improved CTAB method Ⅰ,improved CTAB method Ⅱ,improved CTAB method Ⅲ and improved SDS method.[Result] The four extraction methods could all be successfully used for extracting the genomic DNA from Clematis fasciculiflora Franch.The purity of genomic DNA was the highest using improved CTAB method Ⅰ,with the longest extracting time;while the concentration of genomic DNA was the maximum using the improved SDS method,with the shortest extracting time and relatively low purity;the extracting time of improved CTAB method Ⅲ was the shortest.[Conclusion] This study had established the optimal extraction method for extracting the genomic DNA from Clematis fasciculiflora Franch and supported for the further research using molecular biological methods.
基金Supported by Chongqing Natural Science Foundation~~
文摘[ Objective] The aim was to study method for extracting DNA from pteridophyta and provide basis for further study on genetic diversity and taxonomy. [ Method] By changing the dosage of reagent and operating method, CTAB method for extracting DNA was improved. [ Result] The results showed that the improved CTAB method could extract high-quality DNA from pteridophyta. [ Conclusion] The study improved method for extracting DNA from pteridophyta.
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 ＇
基金Supported by National 863 Program of China(2006AA10A113)Natural Science foundation of Zhejiang Province(Y306097)~~
文摘[Objective] The research aimed to isolate flanking sequences adjacent to the transgenic T-DNA in Brassica napus by an improved inverse PCR method.[Method] Using single clone of transgenic FS4 in Brassica napus as the research materials,total DNA was extracted from transgenic Brassica napus by using modified CTAB method.After enzyme digestion and purification,self-joining was made.Two circles of nested PCR and the sequence alignment were carried out.[Result] A fragement with the size of 4.0 kb was amplified ...
基金Supported by the National Key Technology Research and Development Program of the Ministry of Science and Technology of China(2007BAD47B07-02)Special Foundation for Science and Technology of Chongqing(CSTC,2007AB1040)~~
文摘[Objective] The present study aimed to establish a rapid method for the isolation of small amount DNA from citrus.[Method] By using the improved CTBA method,the genomic DNA was extracted respectively from 20,10,5 and 2.5 mg hybrid embryos of citrus,and then the DNA quality was detected and followed by SSR verification.[Result] The method was very simple and rapid,which needed less materials.In addition,the isolated DNA showed good purity with the OD260/OD280 of 1.8-2.1,and could meet the requirement for PCR-based technology,such as SSR,etc..[Conclusion] The method could be used for rapid extraction of small amount of genomic DNA from citrus.
基金This work was supported by Beijing Natural Science Foundation （No. 6082005） and Funding Project for Academic Human Resources Development in Institutions of Higher Learning under the Jurisdiction of Beijing Municipality （No. PXM2007014207）.
文摘An effective method for isolation of total RNA from pulps of mainly deciduous fruit trees was discussed in this paper. Here we shown that total RNA was successfully isolated from pulps of apple, peach and strawberry, but not grape berry via the effective CTAB method modified by ZENG, et al. However, total RNA was isolated from grape berry according to ZENG＇s protocols with a modification including 16% of β-Mercatoethanol and 3% of PVP in extraction buffer, 1/3 （V/V） of 5mol/L potassium acetate （KAC PH4.8） in supernatant. The detection of the isolated total RNA above using ultraviolet spectrophoto-metry, electrophoresis, reverse transcription and RT-PCR shown that the CTAB and its modifications is a preferred method for isolation of high-quality total RNA from fruit pulps.
