金黄色葡萄球菌px-1 DNA芯片的制备及检测

PREPARATION AND TEST OF DNA MICROARRAY OF THE STAPHYLOCOCCUS AUREUS STRAIN PX-1

提取金黄色葡萄球菌px-1总DNA经酶切后,与载体质粒Puc18链接,并转化E.coliDH5α,从而构成金黄色葡萄球菌px-1的基因文库。用PCR方法扩增基因文库,所得PCR产物点于芯片上,从而制备px-1DNA芯片。分别提取不同盐浓度培养的px-1的总RNA,把不同盐浓度培养的px-1的总RNA反转录为DNA,用Cy3,CY5分别标记反转录的DNA。通过杂交检测芯片,并获得杂交图像。

The total DNA was digested with restricted enzyme, ligated with Puc 18 plasmid and transformed into E. coli DHSa,so,the gene bank was constructed. The PCR Production of px - 1 gene bank was spotted on the slide and then the px - 1 DNA microarray was produced. The purified total RNA was reverse transcripted into DNA and the DNA were labbled with cy3 - dCTP or cy5 - dCTP. The microarray was tested by hybridization and image of the slide was gotten.