摘要
目的 :探讨两种菌落PCR方法在抗体库筛选中的应用价值。 方法 :分别以 5个阳性克隆细菌菌落直接悬浮于去离子水中及悬浮于含 0 .1%的TritonX 10 0后经水浴煮沸作为模板 ,进行PCR反应。同时提取质粒DNA ,进行双酶切鉴定。 结果 :5个克隆的菌落PCR及酶切鉴定均检测到目的基因。 结论 :菌落PCR可用于筛选阳性重组克隆。
Objective:To value the application of two bacteria colony-PCR methods in the screening of phage antibody library. Methods:Five positive monoclonal bacterium were respectively suspended in either deionized water or 0.1% Triton X-100, and then boiled to be used as template in PCR. . The DNAs products of PCR were extracted and digested by two enzymes, and then determined by electrophoresis. Results:The inserted genes were detected in all the 5 clones after PCR and enzyme digestion .Conclusion:Bacteria colony-PCR can be used in screening positive recombinant colonies. The bacteria colony-PCR method with bacteria colonies suspended in deionized water is valuable in large scale positive recombinant bacterium screening.
出处
《医学研究生学报》
CAS
2004年第12期1057-1058,1061,共3页
Journal of Medical Postgraduates
基金
国家自然科学基金资助项目 (批准号 :3 0 3 713 99)
关键词
聚合酶链反应
菌落
抗体库
目的基因
PCR
Bacteria colonies
Phage antibody library
Insert genes
