目的获得15个短串联重复(Short tandem repeat,STR)基因座在西藏藏族人群中的基因型及等位片段频率分布,获得相应位点的群体遗传学数据。方法126份EDTA抗凝血样采自西藏藏族地区无血缘关系的藏族个体。Chelex法提取DNA,PCR复合扩...目的获得15个短串联重复(Short tandem repeat,STR)基因座在西藏藏族人群中的基因型及等位片段频率分布,获得相应位点的群体遗传学数据。方法126份EDTA抗凝血样采自西藏藏族地区无血缘关系的藏族个体。Chelex法提取DNA,PCR复合扩增,自动基因分析仪电泳收集电泳结果数据,基因扫描分析软件计算扩增产物片段相对大小,基因分型软件进行样本基因型分型。结果全部样本的每个STR基因座都获得了清晰的基因型分型结果。15个STR基因座的杂合度介于0.627-0.897之间。累计非父排除率和累计个人识别机率为0.999999527和〉0.999999999。结论经一次扩增电泳可获得15个STR基因座的基因型分型结果,累计非父排除率和累计个人识别机率较高,适用于法医学亲权鉴定和个人识别。展开更多
Determining the base sequence of DNA broken site is quite crucial for the study on the cleavage site specificity and mechanism of various natural or synthetic DNA cleavage regents,and on developing novel therapeutic d...Determining the base sequence of DNA broken site is quite crucial for the study on the cleavage site specificity and mechanism of various natural or synthetic DNA cleavage regents,and on developing novel therapeutic drugs targeting at DNA.The most frequently used method depending on chemical reactions of the Maxam-Gilbert procedure,and the late arising methods used by Rui Ren et al.which were based on Sanger’s DNA sequencing strategy,all had some deficiencies,either the pollution of radioactive materials,or really complicated and difficult to operate.In the present paper,a new method for DNA cleavage site sequence determination was developed.The fluorescence FAM-labeled primer was annealed to the DNA fragments,which has been cleaved by restriction enzymes or other regents,and extended along the template sequence.The products then loaded onto the polyacrylamide electrophoresis gel of ABI 377 DNA Sequencer.Data was collected and analyzed by using ABI PRISM Data Collection Software and ABI PRISM Sequencing Analysis Software.It is proved to be a credible and simple new approach to determine the base sequence of DNA broken sites.展开更多
Studies on the bacterial predation rate by rumen protozoa were carried out under laboratory conditions using a technique of fluorescence-labeled bacteria (FLB). Four Xuhuai goats were used in this experiment to obta...Studies on the bacterial predation rate by rumen protozoa were carried out under laboratory conditions using a technique of fluorescence-labeled bacteria (FLB). Four Xuhuai goats were used in this experiment to obtain rumen protozoa and bacteria. Two groups were designed as follows: One group was the whole bacteria which were labeled using fluorescence through removing free bacteria from rumen fluid (WFLB); the other group was the bacteria which were labeled using fluorescence without removing free bacteria from rumen fluid (FLB). The result indicated that the bacterial predation rates of rumen protozoa was 398.4 cells/(cell h) for the group WFLB, 230.4 cells/(cell h) for the group FLB, when the corresponding values expressed as bacteria-N, they were 2.15 pg N/(cell h) for the group WFLB, and 1.24 pg N/(cell h) for the group FLB, respectively. Extrapolating the assimilation quantity of nitrogen by ciliates on bacteria of Xuhuai goat, there were 103.2 mg N/(d capita) for the group WFLB, and 59.5 mg N/(d capita) for the group FLB, respectively. It was estimated that protein losses due to microbial recycling were 0.645 g pro/(d capita) for the group WFLB and 0.372 g pro/(d capita) for the group FLB, respectively. In addition, the fluorescence-labeled technique would be a potential assay for the determination of bacterial predation rate by rumen protozoa.展开更多
文摘目的获得15个短串联重复(Short tandem repeat,STR)基因座在西藏藏族人群中的基因型及等位片段频率分布,获得相应位点的群体遗传学数据。方法126份EDTA抗凝血样采自西藏藏族地区无血缘关系的藏族个体。Chelex法提取DNA,PCR复合扩增,自动基因分析仪电泳收集电泳结果数据,基因扫描分析软件计算扩增产物片段相对大小,基因分型软件进行样本基因型分型。结果全部样本的每个STR基因座都获得了清晰的基因型分型结果。15个STR基因座的杂合度介于0.627-0.897之间。累计非父排除率和累计个人识别机率为0.999999527和〉0.999999999。结论经一次扩增电泳可获得15个STR基因座的基因型分型结果,累计非父排除率和累计个人识别机率较高,适用于法医学亲权鉴定和个人识别。
文摘Determining the base sequence of DNA broken site is quite crucial for the study on the cleavage site specificity and mechanism of various natural or synthetic DNA cleavage regents,and on developing novel therapeutic drugs targeting at DNA.The most frequently used method depending on chemical reactions of the Maxam-Gilbert procedure,and the late arising methods used by Rui Ren et al.which were based on Sanger’s DNA sequencing strategy,all had some deficiencies,either the pollution of radioactive materials,or really complicated and difficult to operate.In the present paper,a new method for DNA cleavage site sequence determination was developed.The fluorescence FAM-labeled primer was annealed to the DNA fragments,which has been cleaved by restriction enzymes or other regents,and extended along the template sequence.The products then loaded onto the polyacrylamide electrophoresis gel of ABI 377 DNA Sequencer.Data was collected and analyzed by using ABI PRISM Data Collection Software and ABI PRISM Sequencing Analysis Software.It is proved to be a credible and simple new approach to determine the base sequence of DNA broken sites.
基金carried out under the financial support from the National Natural Science Foundation of China(30571344).
文摘Studies on the bacterial predation rate by rumen protozoa were carried out under laboratory conditions using a technique of fluorescence-labeled bacteria (FLB). Four Xuhuai goats were used in this experiment to obtain rumen protozoa and bacteria. Two groups were designed as follows: One group was the whole bacteria which were labeled using fluorescence through removing free bacteria from rumen fluid (WFLB); the other group was the bacteria which were labeled using fluorescence without removing free bacteria from rumen fluid (FLB). The result indicated that the bacterial predation rates of rumen protozoa was 398.4 cells/(cell h) for the group WFLB, 230.4 cells/(cell h) for the group FLB, when the corresponding values expressed as bacteria-N, they were 2.15 pg N/(cell h) for the group WFLB, and 1.24 pg N/(cell h) for the group FLB, respectively. Extrapolating the assimilation quantity of nitrogen by ciliates on bacteria of Xuhuai goat, there were 103.2 mg N/(d capita) for the group WFLB, and 59.5 mg N/(d capita) for the group FLB, respectively. It was estimated that protein losses due to microbial recycling were 0.645 g pro/(d capita) for the group WFLB and 0.372 g pro/(d capita) for the group FLB, respectively. In addition, the fluorescence-labeled technique would be a potential assay for the determination of bacterial predation rate by rumen protozoa.