利用限制性显示 (RD PCR)技术快速分离HIV 1基因片段制备DNA芯片探针 .以Sau3AⅠ酶切HIV基因 ,得到许多大小适合芯片的限制性酶切片段 .然后在片段两端接上接头 ,根据酶切位点、接头的序列设计通用引物 .在该通用引物的 3′端分别延伸一个碱基后 ,通过引物间的两两组合 ,将PCR反应分成 10个亚组 .纯化各组PCR产物 ,克隆到T载体上 .阳性克隆经鉴定、扩大培养后提取质粒 .以质粒为模板扩增靶片段并进行序列分析 .每个亚型得到了十几个 10 0～ 10 0 0bp的HIV基因片段 .研究表明 ,RD PCR技术是一种有效的快速制备基因芯片探针的方法 The HIV 1 gene fragment was rapidly isolated by restriction-based display (RD PCR) technology to prepare DNA chip probes. The HIV gene was digested with Sau3A I to obtain a number of restriction enzyme fragments of suitable size for the chip. Then the linker was ligated to both ends of the fragment, Enzyme digestion site, linker universal primer design universal primer in the 3 ’end of each extension of a base, through a pair of combinations between the two primers, the PCR reaction is divided into 10 subgroups PCR products were purified, And cloned into T vector.The positive clones were identified and expanded after the extraction of the plasmid.The plasmid was used as a template to amplify the target fragment and sequence analysis.Each subtype got a dozen of 10 ~ 100bp HIV gene fragment. Research shows that RD PCR technology is an effective and rapid method of preparing gene chip probes