目的研究1种高纯度牛凝血酶的制备工艺。方法采用不同终浓度的Ba^2＋吸附、硫酸铵分段盐析从牛血浆中提取凝血酶原,用Ca^2＋激活,活化的凝血酶用CM-Sepharose Fast Flow离子交换柱纯化。结果用冷冻1周血浆制备的凝血酶粗品产量、比活明...目的研究1种高纯度牛凝血酶的制备工艺。方法采用不同终浓度的Ba^2＋吸附、硫酸铵分段盐析从牛血浆中提取凝血酶原,用Ca^2＋激活,活化的凝血酶用CM-Sepharose Fast Flow离子交换柱纯化。结果用冷冻1周血浆制备的凝血酶粗品产量、比活明显高于来自新鲜血浆的粗品;Ba^2＋终浓度在0.107 mol/L以上时凝血酶的产量较高;凝血酶原的最佳激活条件为：Ca^2＋终浓度0.05 mol/L,37℃,30～60 min;CM-Sepharose Fast Flow柱制备的凝血酶比活达到2 000 U/mg以上。结论该工艺适用于高纯度凝血酶的制备。展开更多
The DNATyper^(TM)21 kit has good species specificity,typing accuracy,and locus amplification balance,which can be used for forensic genetic analysis such as individual identification and paternity testing.To improve t...The DNATyper^(TM)21 kit has good species specificity,typing accuracy,and locus amplification balance,which can be used for forensic genetic analysis such as individual identification and paternity testing.To improve the anti-inhibitor ability of this kit,the addition of bovine serum albumin(BSA),bovine thrombin(BT),FastStart Taq DNA Polymerase,TaKaRa Ex Taq Hot Start Version(Ex Taq HS),MyFiTM DNA Polymerase,and Klentaq DNA Polymerase(Klentaq)as PCR enhancers to the PCR reaction system was explored in the presence of different concentrations of inhibitors,such as indigo,humic acid(HA),hemoglobin,heme,and melanin.The results revealed that BSA exhibited high efficiency in improving the detection rate of STR loci;BT only helped to overcome the typing inhibition caused by indigo,melanin,HA,and heme;Four types of DNA polymerase had different anti-inhibitor effects on different inhibitors,among which Ex Taq HS was more effective than the other three DNA polymerases,but displayed comparatively lower resistance to hemoglobin than BSA and BT.This study provides basic data for further optimization of the kit through comprehensive analysis and facilitates its application in daily forensic investigations.展开更多
文摘目的研究1种高纯度牛凝血酶的制备工艺。方法采用不同终浓度的Ba^2＋吸附、硫酸铵分段盐析从牛血浆中提取凝血酶原,用Ca^2＋激活,活化的凝血酶用CM-Sepharose Fast Flow离子交换柱纯化。结果用冷冻1周血浆制备的凝血酶粗品产量、比活明显高于来自新鲜血浆的粗品;Ba^2＋终浓度在0.107 mol/L以上时凝血酶的产量较高;凝血酶原的最佳激活条件为：Ca^2＋终浓度0.05 mol/L,37℃,30～60 min;CM-Sepharose Fast Flow柱制备的凝血酶比活达到2 000 U/mg以上。结论该工艺适用于高纯度凝血酶的制备。
文摘The DNATyper^(TM)21 kit has good species specificity,typing accuracy,and locus amplification balance,which can be used for forensic genetic analysis such as individual identification and paternity testing.To improve the anti-inhibitor ability of this kit,the addition of bovine serum albumin(BSA),bovine thrombin(BT),FastStart Taq DNA Polymerase,TaKaRa Ex Taq Hot Start Version(Ex Taq HS),MyFiTM DNA Polymerase,and Klentaq DNA Polymerase(Klentaq)as PCR enhancers to the PCR reaction system was explored in the presence of different concentrations of inhibitors,such as indigo,humic acid(HA),hemoglobin,heme,and melanin.The results revealed that BSA exhibited high efficiency in improving the detection rate of STR loci;BT only helped to overcome the typing inhibition caused by indigo,melanin,HA,and heme;Four types of DNA polymerase had different anti-inhibitor effects on different inhibitors,among which Ex Taq HS was more effective than the other three DNA polymerases,but displayed comparatively lower resistance to hemoglobin than BSA and BT.This study provides basic data for further optimization of the kit through comprehensive analysis and facilitates its application in daily forensic investigations.
