[Objective ] The study aimed to construct the AFLP molecular marking system in Mangifera indica. [ Method ] Four varieties of Mangifera indica were used to explore new ways for high-quality DNA, and AFLP analysis of 3...[Objective ] The study aimed to construct the AFLP molecular marking system in Mangifera indica. [ Method ] Four varieties of Mangifera indica were used to explore new ways for high-quality DNA, and AFLP analysis of 31 varieties of Mangifera indica was carried out to detect the varietal genetic diversity. [ Result] 14 pairs of primers with stronger polymorphism, better banding patterns and higher resolution were screened out from 64 pairs of selective amplification primers. Then they were used to analyse the fingerprint of 31 varieties of Mangifera indica, the results showed that the ratio of polymorphic bands amplificated by the 14 pairs of primers reached 97% in 31 varieties of Mangifera.[ Conclusion] It was suggested that AFLP was suitable for detecting the polymorphism of Mangifera indica resources.展开更多
[Objective] This study aimed to establish an AFLP reaction system for spinach. [Method] Modified CTAB method was used for DNA extraction from six varieties of spinach. Experimental conditions in enzyme digestion and l...[Objective] This study aimed to establish an AFLP reaction system for spinach. [Method] Modified CTAB method was used for DNA extraction from six varieties of spinach. Experimental conditions in enzyme digestion and ligation, pre-amplification and selective amplification in the AFLP reaction system were analyzed and optimized. [Result] The results showed that： （1） enzyme digestion had better effects in the PCR instrument than in a water bath and it was not sensitive to the digestion time and DNA concentrations; （2） a total volume of 20 μl, containing 1.2 μl of Mg2＋ （25 mmol/L）, 1.6 μl of dNTPs （2.5 mmol/L） and 0.2 μl of Taq-polymerase （5 U/μl） was the optimal condition for pre-amplification and selective amplification reactions. [Conclusion] The AFLP system established in this study was stable and efficient, which provided a theoretical basis for AFLP molecular marker analysis of genetic relationship between spinach varieties and genetic breeding.展开更多
[Objective] The obtained clear AFLP fingerprint of Chinese cabbage provided basis for studies on the molecular markers of Chinese cabbage cultivars and the phylogenetic relationship among Chinese cabbage cultivars. [M...[Objective] The obtained clear AFLP fingerprint of Chinese cabbage provided basis for studies on the molecular markers of Chinese cabbage cultivars and the phylogenetic relationship among Chinese cabbage cultivars. [Method] With the test materials of leaves of Chinese cabbages, the high-quality total DNA from leaves of Chinese cabbages was extracted by the modified CTAB method. DNA restriction-ligase reaction, pre-amplification and selective amplification were optimized, and the AFLP silver-staining reaction system for Chinese cabbage was established. [Result] The quality of DNA template influenced restriction enzyme digestion and the subsequent ligase amplification reaction, while the modified CTAB extraction method could be used in AFLP analysis of Chinese cabbage to obtain a clear AFLP fingerprint. The optimum conditions for restriction enzyme digestion of genomic DNA from Chinese cabbage were as follows: 150 g DNA template, 12.5 μl reaction volume, 1.25 U Eco R Ⅰ, 1.25 U Mse Ⅰ and 5×Reaction Buffer with 4 h at 37 ℃. The ligation reaction with 2.5 h at 20 ℃ was the optimum condition. Six pairs of primers including E-AAC/M-CAG, E-AAG/M-CAC, E-ACA/M-CTG, E-ACT/M-CAC, E-ACT/M-CTT and E-ACT/M-CTC all had its own stable and clear patterns. [Conclusion] With abundant bands and high polymorphism, AFLP selective amplification is an efficient molecular marker for genomic polymorphism of Chinese cabbage.展开更多
The optimized AFLP analysis system was established for Alnus. A. glutinosa was used to comparatively study and analyze key factors influencing the results of AFLP, including quality and concentration of extracted DNA,...The optimized AFLP analysis system was established for Alnus. A. glutinosa was used to comparatively study and analyze key factors influencing the results of AFLP, including quality and concentration of extracted DNA, enzyme digestion time and amplification system that influenced the results of the AFLP. The results suggested an amount of DNA template of 300 ng; reaction time for enzyme digestion of 5 h; ligation time of 16 h; a 10 times dilution of ligation products for pre-amplification ; and a 20 times dilution of pre-amplification products for selective amplification. The AFLP-PCR reaction system （20 μl） for Alnus was determined as follows ： DNA template 300 ng, 10 x Taq DNA polymerase Buffer （ containing Mg2＋ ） 2.0μl, dNTPs 0. 3 μl, primers 1.0 μl, Taq DNA polymerase 0. 6 U, and ddH2O in balance. Seven pairs of primer combinations were selected with many clear bands and high polymorphism. By using the AFLP reaction system, primers suitable for Alnus were obtained. Those results provided certain basis for further molecular studies on Alnus using AFLP markers.展开更多
基金Supported by Key Projects of Basic Platform of the National Science and Technology (2005DKA21005 )National Natural Science Foundation-funded Projects (30260086)~~
文摘[Objective ] The study aimed to construct the AFLP molecular marking system in Mangifera indica. [ Method ] Four varieties of Mangifera indica were used to explore new ways for high-quality DNA, and AFLP analysis of 31 varieties of Mangifera indica was carried out to detect the varietal genetic diversity. [ Result] 14 pairs of primers with stronger polymorphism, better banding patterns and higher resolution were screened out from 64 pairs of selective amplification primers. Then they were used to analyse the fingerprint of 31 varieties of Mangifera indica, the results showed that the ratio of polymorphic bands amplificated by the 14 pairs of primers reached 97% in 31 varieties of Mangifera.[ Conclusion] It was suggested that AFLP was suitable for detecting the polymorphism of Mangifera indica resources.
基金Supported by Beijing New Star Science and Technology Program(2008B37)Scientific and Technological Innovation Capacity Building Project of Beijing Academy of Agriculture and Forestry Sciences(KJCX201101010)~~
文摘[Objective] This study aimed to establish an AFLP reaction system for spinach. [Method] Modified CTAB method was used for DNA extraction from six varieties of spinach. Experimental conditions in enzyme digestion and ligation, pre-amplification and selective amplification in the AFLP reaction system were analyzed and optimized. [Result] The results showed that： （1） enzyme digestion had better effects in the PCR instrument than in a water bath and it was not sensitive to the digestion time and DNA concentrations; （2） a total volume of 20 μl, containing 1.2 μl of Mg2＋ （25 mmol/L）, 1.6 μl of dNTPs （2.5 mmol/L） and 0.2 μl of Taq-polymerase （5 U/μl） was the optimal condition for pre-amplification and selective amplification reactions. [Conclusion] The AFLP system established in this study was stable and efficient, which provided a theoretical basis for AFLP molecular marker analysis of genetic relationship between spinach varieties and genetic breeding.
基金Supported by Construction and Industrialization Development of Beijing Vegetable Germplasm Modified Center(H022020130130)the Great Project of Natural Science Foundation of Beijing (5050001)~~
文摘[Objective] The obtained clear AFLP fingerprint of Chinese cabbage provided basis for studies on the molecular markers of Chinese cabbage cultivars and the phylogenetic relationship among Chinese cabbage cultivars. [Method] With the test materials of leaves of Chinese cabbages, the high-quality total DNA from leaves of Chinese cabbages was extracted by the modified CTAB method. DNA restriction-ligase reaction, pre-amplification and selective amplification were optimized, and the AFLP silver-staining reaction system for Chinese cabbage was established. [Result] The quality of DNA template influenced restriction enzyme digestion and the subsequent ligase amplification reaction, while the modified CTAB extraction method could be used in AFLP analysis of Chinese cabbage to obtain a clear AFLP fingerprint. The optimum conditions for restriction enzyme digestion of genomic DNA from Chinese cabbage were as follows: 150 g DNA template, 12.5 μl reaction volume, 1.25 U Eco R Ⅰ, 1.25 U Mse Ⅰ and 5×Reaction Buffer with 4 h at 37 ℃. The ligation reaction with 2.5 h at 20 ℃ was the optimum condition. Six pairs of primers including E-AAC/M-CAG, E-AAG/M-CAC, E-ACA/M-CTG, E-ACT/M-CAC, E-ACT/M-CTT and E-ACT/M-CTC all had its own stable and clear patterns. [Conclusion] With abundant bands and high polymorphism, AFLP selective amplification is an efficient molecular marker for genomic polymorphism of Chinese cabbage.
基金Supported by National Science&Technology Support Program during the"12th Five-Year"(2012BAD01B0604)Central Public-interest Scientific Institution Basal Research Fund(RISF2013010)
文摘The optimized AFLP analysis system was established for Alnus. A. glutinosa was used to comparatively study and analyze key factors influencing the results of AFLP, including quality and concentration of extracted DNA, enzyme digestion time and amplification system that influenced the results of the AFLP. The results suggested an amount of DNA template of 300 ng; reaction time for enzyme digestion of 5 h; ligation time of 16 h; a 10 times dilution of ligation products for pre-amplification ; and a 20 times dilution of pre-amplification products for selective amplification. The AFLP-PCR reaction system （20 μl） for Alnus was determined as follows ： DNA template 300 ng, 10 x Taq DNA polymerase Buffer （ containing Mg2＋ ） 2.0μl, dNTPs 0. 3 μl, primers 1.0 μl, Taq DNA polymerase 0. 6 U, and ddH2O in balance. Seven pairs of primer combinations were selected with many clear bands and high polymorphism. By using the AFLP reaction system, primers suitable for Alnus were obtained. Those results provided certain basis for further molecular studies on Alnus using AFLP markers.
