作　　者： ; ; (郑文岭）;

机构地区： 南方医科大学分子生物学研究所

出　　处： 《生物技术通报》 2005年第1期44-46,共3页

摘　　要： 对测序中的模板、引物、测序反应条件及测序反应纯化方法和仪器操作等进行研究。结果显示测序模板的纯度影响测序的质量 ,浓度对测序的长度有影响。引物设计时除符合一般设计原则外 ,Tm值最好在5 0℃～ 6 0℃之间 ,且无成串的G、C。改变变性、退火、延伸的时间和温度对特殊DNA模板的序列测定有较好的效果。测序反应产物的纯化有几种方法 ,以 70 %乙醇沉淀法最经济、方便。因此模板的纯度和浓度对测序成功与否起决定作用。最佳反应条件可降低成本 ,提高测序成功率 ,乙醇沉淀法是首选的测序反应产物纯化方法。仪器操作熟练、正确与否也会影响测序结果。 The effects of DNA templates,primers,cycle sequencing reaction conditions, purification methods,operation of instrument were comparatively analyzed. The quality of sequencing was influenced by the purity of templates,the length of sequencing was influenced by the concentration of templates.The Tm value of primers should be between 50℃～60℃ and there shouldn’t be too much G+C in the primers.The alteration of the temperature and time of the denaturation,annealing,extension facilitated sequencing of some of the DNA templates.The purity and concentration of DNA templates were closely related to the quality sequence data.The optimized reaction conditions could decrease the cost of sequencing and improve the ratio of success.The ethanol precipitation was the optimized method of products purification of the sequencing reactions. Instrument operation could also influence the sequencing results.

关 键 词： 测序 常见 乙醇沉淀法 影响 改变 成功率 序列测定 浓度 纯度 引物

领　　域： [生物学] [农业科学] [农业科学] [农业科学]