Construction and characterization of a BAC library from a gynogenetic channel catfish <it>Ictalurus punctatus</it>
2003
Wilson Melanie | Wolters William R | Miller Norman W | Katagiri Takayuki | Quiniou Sylvie MA | Waldbieser Geoffrey C
<p>Abstract</p> <p>A bacterial artificial chromosome (BAC) library was constructed by cloning <it>Hind</it>III-digested high molecular weight DNA from a gynogenetic channel catfish, <it>Ictalurus punctatus</it>, into the vector pBeloBAC11. Approximately 53 500 clones were arrayed in 384-well plates and stored at -80°C (CCBL1), while clones from a smaller insert size fraction were stored at -80°C without arraying (CCBL2). Pulsed-field gel electrophoresis of 100 clones after <it>Not</it>I digestion revealed an average insert size of 165 kb for CCBL1 and 113 kb for CCBL2. Further characterization of CCBL1 demonstrated that 10% of the clones did not contain an insert. CCBL1 provides a 7.2-fold coverage of the channel catfish haploid genome. PCR-based screening demonstrated that 68 out of 74 unique loci were present in the library. This represents a 92% chance to find a unique sequence. These libraries will be useful for physical mapping of the channel catfish genome, and identification of genes controlling major traits in this economically important species.</p>
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