【摘要】 目的　建立人胎儿骨骼和关节快速扩增 c DNA末端 (rapid amplification of c DNA ends,RACE c DNA)文库 ,为分离骨骼和关节发育相关基因奠定基础。方法　采用改进的异硫氰酸胍 -酚 -氯仿 -异戊醇一步法提取骨骼和关节总 RNA,用 Ta Ka Ra公司生产的 c DNA合成试剂盒合成平末端的双链 c DNA,然后与衔接子连接。再用位于双链 c DNA末端的通用引物扩增全部 c DNA。结果　建立了从骨骼和关节构建 RACE c DNA文库的方法 ,并用该方法成功地构建了人胎儿骨骼和关节 RACE c DNA文库。结论　所构建的的利用少量总 RNA构建 RACE c DNA文库方法切实可行 ,所构建的文库适用于用 RACE方法从中分离骨骼和关节发育相关基因更多还原

【Abstract】 Objective To construct rapid amplification cDNA ends(RACE) cDNA libraries from human fetal bone and joint and provide resources for isolation of bone and joint specific development related genes.Methods Total RNA of bone and joint were extracted with the modified single step method of RNA isolation by acid guanidinium thiocyanate phenol chloroform extraction. The double stranded end blunted cDNA were synthesized using TaKaRa’s cDNA synthesis kit and ligated to cassette adaptors. All of the cDNA molecules were amplified by a pair of common primers.Results A protocol for RACE cDNA library construction from bone and joint was established and two RACE cDNA libraries from human fetal bone and joint were successfully constructed.Conclusion The protocol of RACE cDNA library construction from limited materials proved to be simple and efficient and the library was suitable for RACE to isolate tissue specific genes.更多还原