DNA测序中常见影响因素的研究-生物技术通报.PDFVIP

生物技术通报 BIOTECHNOLOG Y B ULLETIN 2005 1 DNA 1 1 2 宋艳斌 马文丽 郑文岭 1 2 ( , 510515; , 5 100 10) : 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 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 50 ~ 60 之间, 且无成串的GC 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 : DNA 循环测序 影响因素 The Influence of Some Factors on DNA Sequencing 1 1 2 Song Yanbin Ma Wen i Zheng Wen ing 1 ( Dep artment of Biochemistry , the First Military Medical University , Guangzhou 5105 15; 2 Institute of Molecular Oncology, iu Hua Qiao Hosp ital , Guangzhou 5100 10) Abstract : The effects of DNA temp ates, primers, cyc e sequencing reaction conditions, purification methods, operation of in strument w ere comparative y ana yzed . The qua ity of sequencing w as inf uenced by the purity of temp ates, the ength of sequenc ing w as inf uenced by the concentration of temp ates . The Tm va ue of primers shou d be betw een 50 ~ 60 and there shou dn t be too much G+ C in the primers . The a teration of the temperature and time of the denaturation, annea ing, extension f aci itated sequencing of some of the DNA temp ates . The purity and concentration of DNA temp ates w ere c ose y re ated to the qua ity se quence data.The optimized reaction conditions cou d decrease the