The ISSR method was developed to analyze the genetic diversity of 20 wild Wusuli raccoon dog from Nenjiang district. The results showed that there were significant genetic diversity and high polymorphism among individ...The ISSR method was developed to analyze the genetic diversity of 20 wild Wusuli raccoon dog from Nenjiang district. The results showed that there were significant genetic diversity and high polymorphism among individuals of the wild Wusuli raccoon dog. A total of 41 DNA bands were amplified by 9 primers, and 37 of them were polymorphism, the proportion of polymorphism was 90.24%, 2-8 polymorphism bands could be amplified by each primer, and 4.56 on average, the length of the product was 200-2 000 bp.展开更多
[Objective] This study aimed to investigate a reliable method for DNA ex- traction from Wusuli raccoon dog＇s hair. [Method] Several DNA extraction methods were used to extract DNA from Wusuli raccoon dog hair, includ...[Objective] This study aimed to investigate a reliable method for DNA ex- traction from Wusuli raccoon dog＇s hair. [Method] Several DNA extraction methods were used to extract DNA from Wusuli raccoon dog hair, including Chelex-100 method, PCR buffer method, organic phenol-chloroform method and centrifugal col- umn type kit method. The extracted DNA was analyzed by using PCR amplification and electrophoresis to compare these four DNA extraction methods. [Result] Accord- ing to the results of spectrophotometer detection and gel electrophoresis, nucleic acid extracted by Chetex-100 method had proteins and other impurities; nucleic acid ex- tracted by PCR buffer method was low in concentration; however, DNA extracted by organic phenol-chloroform method and centrifugal column type kit was high in con- centration with no impurity band. [Conclusion] This study had laid the strong founda- tion of scientific theory to further explore the efficient and simple method for extracting DNA from Wusuli raccoon dog hair follicle.展开更多
目的建立一种分离纯化乌苏里蝮蛇血凝酶(WH)的方法并为其制定质量标准。方法先后采用分子筛层析法、离子交换色谱法、亲和层析法从乌苏里蝮蛇毒中分离纯化得到一种具有凝血活性的酶成分,并采用SDS-聚丙烯酰胺凝胶电泳法、RP-HPLC法测定...目的建立一种分离纯化乌苏里蝮蛇血凝酶(WH)的方法并为其制定质量标准。方法先后采用分子筛层析法、离子交换色谱法、亲和层析法从乌苏里蝮蛇毒中分离纯化得到一种具有凝血活性的酶成分,并采用SDS-聚丙烯酰胺凝胶电泳法、RP-HPLC法测定其纯度,HPSEC法测定其分子量,IEF法测定其等电点,Lowry法测定其蛋白浓度,并用标准人血浆法测定该酶的比活力。结果从乌苏里蝮蛇蛇毒中分离纯化了一种凝血酶成分,SDS-Page显示为一条带、测得分子量约为34 k D,RP-HPLC得到单一的色谱峰,HPSEC法测得该酶分子量为34.7 k D,等电点为5.25,此酶具有体外凝血活性,比活力为5.40μg/U。结论该方法可用于WH的分离纯化,制定的质量标准可用于控制该血凝酶的质量。展开更多
文摘The ISSR method was developed to analyze the genetic diversity of 20 wild Wusuli raccoon dog from Nenjiang district. The results showed that there were significant genetic diversity and high polymorphism among individuals of the wild Wusuli raccoon dog. A total of 41 DNA bands were amplified by 9 primers, and 37 of them were polymorphism, the proportion of polymorphism was 90.24%, 2-8 polymorphism bands could be amplified by each primer, and 4.56 on average, the length of the product was 200-2 000 bp.
基金Supported by National Natural Science Foundation of China (31072018)~~
文摘[Objective] This study aimed to investigate a reliable method for DNA ex- traction from Wusuli raccoon dog＇s hair. [Method] Several DNA extraction methods were used to extract DNA from Wusuli raccoon dog hair, including Chelex-100 method, PCR buffer method, organic phenol-chloroform method and centrifugal col- umn type kit method. The extracted DNA was analyzed by using PCR amplification and electrophoresis to compare these four DNA extraction methods. [Result] Accord- ing to the results of spectrophotometer detection and gel electrophoresis, nucleic acid extracted by Chetex-100 method had proteins and other impurities; nucleic acid ex- tracted by PCR buffer method was low in concentration; however, DNA extracted by organic phenol-chloroform method and centrifugal column type kit was high in con- centration with no impurity band. [Conclusion] This study had laid the strong founda- tion of scientific theory to further explore the efficient and simple method for extracting DNA from Wusuli raccoon dog hair follicle.
文摘目的建立一种分离纯化乌苏里蝮蛇血凝酶(WH)的方法并为其制定质量标准。方法先后采用分子筛层析法、离子交换色谱法、亲和层析法从乌苏里蝮蛇毒中分离纯化得到一种具有凝血活性的酶成分,并采用SDS-聚丙烯酰胺凝胶电泳法、RP-HPLC法测定其纯度,HPSEC法测定其分子量,IEF法测定其等电点,Lowry法测定其蛋白浓度,并用标准人血浆法测定该酶的比活力。结果从乌苏里蝮蛇蛇毒中分离纯化了一种凝血酶成分,SDS-Page显示为一条带、测得分子量约为34 k D,RP-HPLC得到单一的色谱峰,HPSEC法测得该酶分子量为34.7 k D,等电点为5.25,此酶具有体外凝血活性,比活力为5.40μg/U。结论该方法可用于WH的分离纯化,制定的质量标准可用于控制该血凝酶的质量。
