Screening for overlapping bacterial artificial chromosome clones by PCR analysis with an arbitrary primer uri icon

abstract

  • In this article, we used PCR analysis with arbitrary primers (AP-PCR) to screen for overlapping bacterial artificial chromosome (BAC) clones and assembly of contigs. A rice BAC library with three genome equivalents was used to prepare pooled BAC DNA, Twenty-two arbitrary primers were used to survey the pooled BAC DNAs and individual BAC DNAs, Each primer identified 1-10 loci, and the average was 4.4 loci. There were 1-5 overlapping clones in each locus, and the average was 2.5 clones, A total of 245 BAC clones were identified as overlapping by AP-PCR and the identities were confirmed by DNA-DNA hybridization. The 245 BAC clones were then assembled into 80 contigs and 17 single-clone loci, The results indicated that PCR analysis with arbitrary primers is a powerful tool in screening for overlapping BAC clones with high accuracy and efficiency, The use of AP-PCR analysis should speed up the construction of physical maps of the plant and animal genomes, as well as the rice genome.

publication date

  • 1998
  • 1998
  • 1998