用聚赖氨酸包被国产载玻片制作DNA芯片

Constructing a DNA microarrary using homemade glass slides coated with poly-L-lysine

用多聚赖氨酸对国产载玻片进行包被处理,在不同的固定条件下,比较其与进口多聚赖氨酸载玻片对大片段DNA和3′端氨基修饰寡核苷酸的固定效果.结果表明,在DNA固定率上,自行包被处理的国产载玻片与进口多聚赖氨酸载玻片没有显著差异;紫外交联对大片段DNA固定率影响显著,但对氨基修饰的寡核苷酸影响不显著;3种点样液中,进口点样液MSS点样效果最好,其次为3×SSC,第三是水.氨基修饰的寡核苷酸可以很好地固定在聚赖氨酸包被的载玻片上.

Two kinds of glass slides coated with poly-L-lysine were used as DNA attachment supports. One was made in our lab using the microscope glass slides made in China as a matrix, the other was a commercial product from Sigma. We immobilized a large fragment of DNA and 3′-amino-modified oligonucleotides on the two kinds of slides under different treatments. DNA attachment rates were investigated and the results indicated that there was no obvious difference between the two kinds of slides. Ultraviolet crosslinking greatly affected the attachment rate of a large fragment of DNA, but did not noticeably affect the attachment rate of the amino-modified oligonucleotides. Among the three spotting solutions, the imported spotting solution MSS was the best one for DNA spotting, 3×SSC was the second, and water was the last. The amino-modified oligonucleotides could be efficiently immobilized on the slide coated with poly-L-lysine.