title: [Construct cosmid libraries by isolating large genomic DNA fragments from Monascus ruber].
abstract: OBJECTIVE
  To isolate large genomic DNA fragments from Monascus ruber for the construction of cosmid libraries.
  
  
  METHODS
  Modified phenol-chloroform method was used to isolate genomic DNA. The isolated genomic DNA was digested by Sau3AI to 40kb fragments on average. Then, the fragments were packaged by Stratagene' s Gigapack III XL packaging extract. A pair of degenerate primers were used to amplify a fragment of PKS (polyketide synthase) gene from this cosmid library.
  
  
  RESULTS
  The average size of genomic DNA isolated by this method was larger than 48 kb, with a concentration of 5 microg/microl. The constructed cosmid libraries had 10 folders coverage of the Monascus spp. genome. A cosmid containing the homologue of PKS gene was obtained by PCR screening.
  
  
  CONCLUSIONS
  This modified method of isolating large fragments genomic DNA of Monascus ruber was efficient and feasible.
authors.affiliation: Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China. anyang@foxmail.com
authors.forename: Yang
authors.initials: Y
authors.lastname: An
authors.forename: Jing
authors.initials: J
authors.lastname: Yang
authors.forename: Xinxin
authors.initials: X
authors.lastname: Xu
authors.forename: Gang
authors.initials: G
authors.lastname: Liu
mesh_terms: D003001:Cloning, Molecular; D003360:Cosmids / Q000235:genetics*; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification*; D015723:Gene Library; D005821:Genetic Techniques; D032741:Monascus / Q000235:genetics*
chemical_list: D004271:DNA, Fungal