60mer寡核苷酸芯片制备及杂交系统的优化  

OPTIMIZATION OF PREPARATION OF 60MER OLIGONUCLEOTIDE MICROARRAY PREPARATION AND HYBRIDIZATION SYSTEM

作　　者：张海燕[1] 马文丽[1] 黄吉城[2] 商涛[1] 廖之君[1] 郭波旋[2] 郑文岭[3] 

机构地区：[1]广州军区广州总医院医学实验科,广州510100 [2]广东出入境检验检疫局检验检疫技术中心 [3]南方医科大学基因工程研究所

出　　处：《现代预防医学》2008年第18期3600-3601,3604,共3页Modern Preventive Medicine

基　　金：国家质量监督检验检疫总局科技计划资助项目(2006IK0168)

摘　　要：[目的]寡核苷酸芯片制备及杂交过程中的影响因素很多,本实验针对其中主要影响因素进行优化,以筛选出最佳的条件,对芯片制备及杂交系统进行优化。[方法]通过改变芯片打印加样量、斑点数和不同探针浓度,挑选出芯片制备的实验条件;采用正交试验法,以信噪比(SNR值)为评估指标,分析芯片杂交的最佳条件。[结果]最佳实验条件为:增加预打印斑点数,探针浓度为1μg/μl,与纯化后的荧光标记样品在50%甲酰胺,10×SSC,0.2%SDS杂交液中42℃杂交4h,洗脱液每次洗脱时间为1min。[结论]此方法适用于60mer寡核苷酸芯片的制备及杂交效率的提高。[Objective ] Many factors could affect the preparation of oligonucleotide tmeroarray and hybridization system, and this study optimized the main influencing factors so as to find the optimal condition and optimize the preparation of oligonucleotide microarray and hybridization system. [ Methods] The experiment condition of preparing oligonucleotide mieroarray was chosen by changing the printing quantity of array, quantity of spot and different density of probe. By orthogonal experiment method and signal-noise ratio as evaluation index, the study analyzed the optimum condition of array hybridization. [Results] The optimum experiment condition were as follows： increasing the quantity of pre-printing spot, the density of probe was 1 μg/μl. Then hybridize with purified fluorescent sample in 50% formamide, 10xSSC, 0.2%SDS at 42% 4h, in wash solution Ⅲ （1 min for each time）. [Conclusion] This method is suitable for preparation of oligonucleotide mieroarray and increasing the efficiency of hybridization system

关 键 词：寡核苷酸芯片 正交试验 优化 

分 类 号：Q78[生物学—分子生物学]