四环素及β-内酰胺耐药性基因芯片的制作及优化  

Optimization the preparation of tetracyclines and β-lactam antibiotic-resistant genes microarrays

作　　者：路浩[1] 王兴龙[1] 李晓艳[1] 郎需龙[1] 王瑞红 张付贤[1] 闫广谋[3] 李研东[3] 

机构地区：[1]军事医学科学院军事兽医研究所,长春130062 [2]河北省动物卫生监督所,石家庄050081 [3]吉林大学畜牧兽医学院,长春130062

出　　处：《中国抗生素杂志》2010年第2期148-151,I0003,共5页Chinese Journal of Antibiotics

基　　金：吉林省科技发展计划资助项目(20050549);军队医药卫生项目(06MB321)

摘　　要：目的优化四环素及-内酰胺耐药性基因芯片制作和杂交条件。方法设置多组实验,逐一改变影响寡核苷酸在玻片表面固定及芯片杂交效率的实验条件,从中优化出芯片实验的最佳条件。结果得出的最佳条件是:探针以30pmol/μL的终浓度溶于50%二甲基亚砜中,在温度为20℃、湿度70%的条件下点制在基片上;荧光探针纯化后使用2×杂交液在42℃湿盒中杂交5h。结论优化出了耐药基因芯片制作和杂交的最佳条件,总结出芯片制作优化的方法。Objective To construct the method for optimizing the preparation of tetracyclines and β-lactam antibiotic-resistant genes microarrays. Methods By experiment of optimizing immobilization and hybridization of microarrays experiments, such as slides, spotting solution dissolving DNA, DNA concentration immobilized on glass surface, spotting temperature and humidity, method of DNA immobilized on glass surface, method of labeling sample, hybridization solution, hybridization temperature, hybridization time, etc. Factors were modified accordingly, and optimal values were selected by comparing those of different groups. Results The efficient hybridization condition is as follows： 30 pmol/μl probes resolved in 50% DMSO and spotting on glass slide at 20℃ and in 70% humidity environment .then hybridized with purified fluorescent sample in 2xhybridization solution and formamide at 42 ℃ for 5 hours. Conclusion The conditions of optimizing immobilization and hybridization of microarrays might be used for the production of microarrays.

关 键 词：基因芯片 制作 优化 

分 类 号：R978.1[医药卫生—药品]