CAJ | 학술논문

目的优化四环素及β-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30 pmol/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5 h.结论优化出了耐药基冈芯片制作和杂交的最佳条件,总结出芯片制作优化的方法.
목적 우화사배소급β-내선알내약성기인심편제작화잡교조건.방법 설치다조실험,축일개변영향과핵감산재파편표면고정급심편잡교효솔적실험조건,종중우화출심편실험적최가조건.결과 득출적최가조건시:탐침이30 pmol/μL적종농도용우50%이갑기아풍중,재온도위20℃、습도70%적조건하점제재기편상;형광탐침순화후사용2×잡교액재42℃습합중잡교5 h.결론 우화출료내약기강심편제작화잡교적최가조건,총결출심편제작우화적방법.
Abstract 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 2×hybridization 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.