Construction Of Goat Genomic Library

A complete genomic library of Xinong saanen goat was successfully constructed using lambda bacteriophage vector for screening milk-protein-encoding gene and construction of knock-in vector. The results is as follows: 1. Goat blood was lysised by sucrose lysis buffer and digested by proteinase K. Genomic DNA prepared for library construction was extracted by phenol-chloroform, OD260/ OD280=1.81, partially digested with Sau3A?. 15-23kb fragment was fractionated. 2. Inserts was ligated with Lambda BlueSTAR BamH? Arms using insert:vector molar ratios 3:1. High cloning efficiency was obtained. 3. Recombinant molecules were packaged in vitro with the Package Extract, then infected E.coli ER1647. Titer of the library is 3.4×10~6 pfu/μg DNA. 4. Plasmid pBlueSTAR-insert subclones are easily obtained by plating ?BlueSTAR phage on a strain(BM25.8) expressing cre recombinase in the presence of carbenicillin. Inserts size were identified by Plasmid NotI digestion. 5. 1.7×10~6 clones were obtained with inserts size ranging from 15-23kb.