新吉细毛羊皮肤组织均一化全长cDNA文库的质量评价

通过对已构建新吉细毛羊皮肤组织的均一化全长cDNA文库进行活化，在此基础上进行EST生物信息学分析研究，期望能发现影响细毛羊羊毛性状的主效基因或相关基因。运用菌落PCR、菌液PCR、质粒快速鉴定3种方法对cDNA文库进行评价，通过测序，在NCBI上BLASTn搜索、比对及运用DNAstar等软件进行分析。初步鉴定的结果：cDNA文库的库容量（滴度）为1．57×10。pfu／mL，菌落PCR、菌液PCR阳性克隆的小片段率在80％～90％，克隆的片段大小在0．6～2．0kb，cDNA在300～500bp，而质粒快速鉴定的在90％，克隆的片段载体带大小3．5～5．0kb。质粒快速鉴定的效果好于其他两种方法，耗时相对短，同时小片段率高；cDNA文库保存完好，符合基因文库的质量要求，有足够大的库容量来保证筛选目的基因。通过对筛选的阳性克隆5’端随机测序，随后获得可读序列169条，经过相关生物信息查询之后，发现其中可能含有完整基因。

Quality Evaluation of Normalized Full-length cDNA Library of Xinji Fine-wool Sheep Skin Tissue

The objective of this paper was to investigate the normalized full-length eDNA library of Xinji fine wool sheep skin tissue, based on which EST bioinformatics were analyzed to expect to find the main effect genes or related genes,and then to evaluate eDNA library by use of three methods including colony PCR,the culture fluid PCR, plasmid fast indentification of eDNA library. By sequencing, the BLASTn search,comparison were analyzed by the use of DNA star software on the NCBI. The preliminary results showed that the storage capacity of the eDNA library was 1.57 X 106 pfu/mL, colong PCR, small fragment rate positively cloned by the culture fluid PCR was 80%- 90%,the size of cloned fragment was 0.6-2.0 kb, eD- NA was 300-500 bp, and plasmid rapid identification was about 90%, with about 3.5- 5.0 kb the cloned fragment size carrier . Ptasmid rapid identification of the effect a little better than the other two methods, relatively short time was cost,while small fragment was slightly higher, cDNA library was pre- served in good condition which to meets the quality requirements of the gene library,a storage capacity is large enough to ensure screening purposes gene. Through the screening positive clones were 57 randomly sequened,then was obtained readable sequence 169 gain,after a bioinformatics inquiry, full gene may be contained in it. Which set the base for further developing related genes with wool traits.