摘要
建立一种简便、快速、特异的制备基因芯片探针的方法.以K 562细胞和正常人淋巴细胞作为消减对象,利用自行建立的消减方法进行消减杂交,结合限制性显示技术,分组扩增差异cD N A,回收K 562细胞特异基因片段,制作基因芯片探针.结果显示,分离到400个K 562特异的基因,片段大小均一,适于制作cD N A芯片.消减杂交技术结合限制性显示技术制备基因芯片探针,具有快速、简便、特异的特点,降低了芯片制作成本,可加速芯片的推广应用.
A fast, easy and specific probe preparation method of microarray was studied. Double-strand cDNA from K562 cells （tester） and normal lymphocyte （driver） were synthesized by reverse transcription respectively. After three times of subtractive hybridization and polymerase chain reaction （PCR） amplification in subgroups with restriction display, the cDNA fragments were isolated by PAGE and secondary amplified to prepare probes of microarray. 400 specific cDNA fragments of K562 cells were obtained as probes of microarray using subtraction hybridization and restriction display. The probes length is comparatively homogeneous and suitable for microarray fabrication. Combination of subtraction hybridization and restriction display technology could provide a fast, easy and specific method for probe-preparation of microarray.
出处
《生命科学研究》
CAS
CSCD
2005年第3期238-241,共4页
Life Science Research
基金
国家自然科学基金资助项目(39880032)
