国家重点基础研究发展规划项目(973)(G1999055903)和中国科学院知识创新工程项目(KSCX-Z-SW-201和KSCX-IOZ-07).
This work was supported by grants from The Special Funds for Major State Basic Research Project (G1999055903) and The Knowledge Innovation Project of The Chinese Academy of Sciences (KSCX-2-SW-201 and KSCX-IOZ-07).
利用根据cap-finder方法建立的全长cDNA合成技术,扩增获得了恒河猴着床点子宫内膜组织表达mRNA的双链cDNA,通过抑制性消减杂交,成功地构建了恒河猴着床点消减文库.随机挑选文库中的阳性克隆,经点杂交证明27%为着床点差异表达的克隆.由此表明抑制性消减杂交结合cap-finder扩增全长cDNA的方法,可以有效地从少量而珍贵的样本中获得高质量的消减文库.
A full-length cDNA amplification technique based on the cap-finder method was used to obtain amplified amount of double strand cDNAs of the endometrial mRNAs at the implantation site from pregnant rhesus monkey. Then a subtractive library of the implantation site was successfully constructed by suppression subtractive hybridization (SSH). Of the randomly selected clones from the library,27% were proved by dot blot analysis to be differentially expressed ones at the implantation site. The data demonstrated that a subtractive library with high quality could be constructed from small amount of rare specimens by the combination of suppression subtractive hybridization(SSH)with full-length cDNA amplification technique based on the cap-finder method.
孙晓阳,谈寅飞,唐爽,王雁玲.用少量样本进行抑制性消减杂交[J].生物化学与生物物理进展,2003,30(6):889-893
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