单细胞cDNA的制备及扩增

Preparation and amplification of cDNAs from a single cell

目的建立微量细胞的基因表达分析系统，解决细胞全基因组表达模式分析和同质细胞材料获取困难之间的矛盾。方法挑取单细胞直接进行逆转录，随后以１个含有ｏｌｉｇｏ（ｄＴ）的通用引物进行序列非依赖性的ｃＤＮＡ扩增并检测其代表性。结果从单个ＨＬ－６０细胞中成功地扩增了全ｃＤＮＡ。结论利用单个或少数细胞制备全ｃＤＮＡ探针，为在高分化的异质性系统中进行差异基因表达分析、建立细胞／组织特异性全基因表达图谱提供了一条合理的途径。

Objective To set up the technique by which gene expression analysis could be done from a few cells so as to resolve the challenging problem between increasingly needs of cellular gene expression pattern analysis and the difficulty of obtaining homogeneous cell materials in some cases.HZMethods Pick up a single cell to do reverse transcription directly, and then amplify cDNA in sequenceindependent manner by using a single primer containing oligo (dT) and test the representation of cDNA. Results Representatively amplified cDNAs from a single HL60 cell. ConclusionZ]Producing whole cellular cDNAs probe from samples as small as a single cell could provide an ideal way to analyze differential gene expression in heterogeneous highly differentiating system and to establish cell type/tissue type specific gene expression profiles.