›› 2007, Vol. 34 ›› Issue (10): 53-55.
• 生物技术 • Previous Articles Next Articles
LI Bing1,YAN Shouqing1,SUN Jinhai1,2
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Abstract: Genomic DNA prepared for library construction was digested by proteinase K and extracted by phenol. The DNA was randomly sheared and endrepaired by T4 DNA Polymerase and T4 Polynucleotide Kinase.25~40 kb fragment was recovered by 1% agarose gel electrophoresis and ligated with pCC1FOS Vector. Recombinant molecules were packaged in vitro with the Phage Packaging Extract, then transformedinto E coli EPI300. A fosmid genomic library of domestic pig was successfully constructed with the titer of 8.50×109 CFU/ml in which the insert fragment size was about 25 kb.Positive clones were achieved by in stitu hybridization with MC1satellite DNA as probe.It was indicated that the library was of high quality which can be used to screen new satellite DNA in domestic pig.
Key words: domestic pig; fosmid library; satellite DNA
CLC Number:
Q785
LI Bing;YAN Shouqing;SUN Jinhai; . Construction of Fosmid Library in Domestic Pig [J]. , 2007, 34(10): 53-55.
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