Method for cloning restriction fragments containing the termini of BAC inserts.

@article{Plyler2000MethodFC,
  title={Method for cloning restriction fragments containing the termini of BAC inserts.},
  author={Tammy R Plyler and C. Eduardo Vallejos},
  journal={BioTechniques},
  year={2000},
  volume={28 5},
  pages={
          1012-6, 1018
        }
}
We have developed a method to isolate the termini of BAC clones. The method is based on the two unique NotI sites located approximately 300 bp on either side of the EcoRI cloning site of the BAC vector pECS-BAC4. Our strategy includes the following steps: (i) generation of Southern blots with BAC clones digested with NotI and a second restriction enzyme; (ii) identification of the termini attached to the NotI/EcoRI fragment of the BAC vector via hybridization with a probe derived from sequences… CONTINUE READING