小鼠胚泡着床期子宫内膜LongSAGE基因文库构建

Construction of a LongSAGE Library of Mouse Endometrium during Implantation

按照LongSAGE文库构建原理,先后经子宫内膜总RNA提取、cDNA合成、130bp双标签的产生、PCR扩增、34bp双标签形成、连接成串联体、与载体连接、转化大肠杆菌等相关步骤构建小鼠胚泡着床期子宫内膜LongSAGE基因文库。经证实所构建文库的阳性克隆率达100%,克隆中插入的串联体长度介于400￣500bp之间,成功构建了小鼠胚泡着床期子宫内膜LongSAGE基因文库。

According to the principle of LongSAGE library construction, we have constrcted the LongSAGE library through several procedures, such as total RNA isolation from mouse endometrium, cDNA synthesis, 130 bp ditag formation, amplification of ditags, isolation of the 34 bp ditags, concatemer formation, cloning the concatemers into pZErO- 1, transformation. It is verified that the positive ratio of constructed LongSAGE library is 100% and the length of inserting fragments is more than 400 bp. These results suggests that the LongSAGE library of mouse endometrium during implantation is successfully generated.