摘要
[目的]比较制备放线菌基因组DNA PCR模板的两种方法液氮研磨法及TE煮沸法。[方法]以从河西走廊盐碱土壤中分离鉴定的8属16株菌为材料,包括糖丝菌属(Saccharothrix)Ⅳ23-3-4、类诺卡氏菌属(Nocardioides)Ⅴ22-5-1、原小单孢菌属(Promicromonospora)Ⅸ5-4-3、小单孢菌属(Micromonospora)Ⅱ23-4-1、放线多孢菌属(Actinopolyspora)Ⅳ8-2-5、拟诺卡氏菌属(Nocardiopsis)Ⅱ8-4-3、链单孢菌属(Streptomonospora)DA01305、链霉菌属金色类群DA03401、链霉菌属金色类群DA01408、链霉菌属蓝色类群DA09140、链霉菌属金色类群DA01308、链霉菌属灰红紫类群DA05213、链霉菌属蓝色类群DA11417、链霉菌属粉红孢类群DA04401、链霉菌属灰红紫类群DA04402、链霉菌属球孢类群DA04307。对两种方法制备的PCR模板进行16S rDNA的扩增,PCR产物进行电泳检测。[结果]两种方法均能得到比较清晰的目的条带,但TE煮沸法简化了放线菌基因组DNA PCR模板制备的过程,可用于高通量放线菌PCR模板的快速制备。[结论]该研究为大批量菌株的快速鉴别和系统分类提供了有效的方法。
[ Objective] The aim was to compare the two preparation methods of PCR templates of genomic Dna from Actinobaceteria. [ Method ] 16 strains of 8 genera separated and identified from saline soil of Langfang, Hexi, including Saccharothrix IV23-3-4, Nocardioides V22-5-1, Promicromonospora 1X5-4-3, Micromortospora 1123-4-1, Actinopolyspora IV8-2-5, Nocardiopsis ]I8-4-3, Streptomonospora DA01305, Streptomyces golden groups DA03401, Streptomyces golden groups DA01408, Streptomyces blue groups DA09140, Streptomyces golden groups DA01308, Streptomyces ash, red and purple groups DA05213, Streptomyces blue groups DA11417, Streptomyces pink spore groups DA04401, Streptomyces ash, red and purple groups DA04402, Streptomyces ball spore groups DA04307 o The liquid nitrogen grinding method and the TE boiling method was used to treat Actinobaceteria in order to explore the rapid preparation method of PCR template, and the PCR template was~ amplified of 16S rDNA, then PCR product was detected by electrophoresis. [ Method] The results showed the two methods both got the clear^0bje^ive band, while the TE boiling method simplified the PCR template preparation process of DNA from Actinobacete.ria, so could be used to l^rephte the PCR tem- plate of high-throughput Actinobaceteria rapidly. [ Conclusion ] The study provides an effective method for repaid identification and systematic classification of large quantities of strains.
出处
《安徽农业科学》
CAS
2013年第23期9569-9570,9574,共3页
Journal of Anhui Agricultural Sciences
基金
国家自然科学基金项目(31260134)
关键词
放线菌
液氮研磨法
TE煮沸法
PCR模板
Actinobaceteria
Liquid nitrogen grinding method
TE boiling method
PCR template
