【摘要】 目的:研究DNA循环测序过程中一些影响因素及其序列图谱。方法:对测序中模板、引物、测序反应条件和体系及其产物的纯化等因素进行比较研究。结果:模板因素是测序成败的主要原因,模板的质量直接影响着测序图谱;引物失效,与模板匹配不完全,Tm值过低等因素也能引起测序失败;利用乙醇沉淀纯化测序反应产物、效果好且简单、经济。结论: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。仪器操作熟练、正确与否也会影响测序结果。更多还原

【Abstract】 Objective:To study the influence of some factors on DNA cyclesequencing by CEQ8800 genetic analytical system.Methods:The effects of DNA templates、primers、cycle sequencing reac-tions as well as purification methods were comparatively analyzed.Results:Sequencing failure was caused mainly by DNA template.The ineffective primers could cause sequencing failure.Possible explanations include primer mismatches and low Tm value.Conclusion:The quality of template DNA can have a major impact on the quality of the DNA sequencing.The optimized reaction conditions can decrease the cost of sequencing and improve the ratio of success.The ethanol precipitation is the optimized method of products purification of the sequencing reactions.Instrument operation can also influence the sequencing results.更多还原