三种寡核苷酸芯片片基的比较

A Robust and Low-cost Substrate for Oligonucleotide Microarray Compared with CMT-GAPS^(TM) Aminosilane Slide and Microscope Slide

将荧光标记的寡核苷酸打印在Corning、多聚赖氨酸包被的DAKO玻片和多聚赖氨酸处理的一种显微镜载玻片上,并按常规方法进行洗脱,通过GenePix4100A扫描并以Genepix6.0软件分析点的大小、荧光强度和背景荧光强度来衡量3种片基的均一性和固定效率,并通过不同浓度梯度的60bp寡核苷酸探针的杂交实验来衡量片基对杂交效率的影响.结果显示,3种片基的均一性都较好,而多聚赖氨酸包被的DAKO和Corn-ing芯片的固定效率和杂交效率优于国产芯片.

Oligonucleotide microarray was pr inted on three different kinds of substrates UltraGAPS TM aminosi-lane slide,poly-L-lysine-coated s lide DAKO and poly-L-lysine modifie d microscope slide.These slides were hybridized with the corresponding Cy3labeled oligonucleotide probe.F ollowing standard washing procedur es,the slides were scanned by GenePix 4100A Scanner and spot sizes.Fluoresc ent intensities and background intensities of three kinds of substr ates were analyzed by Genepix6.0software to evaluate the appropriateness of the substrates.Influences of the su bstrates on hybridization efficien cy were evaluated after Cy3-labeled DNA samples of green fluorescent protein were hybridized to the correspondi ng 60mer oligonucleotide probes.Go od homogeneities were observed on all t he substrates,however,slide Corni ng or DAKO had more stable immobi-lization and showed higher hybridiz ation efficiency than microscope slide.