Preparing Nested-Deletion Template DNA for Field Inversion Gel Electrophoresis Analyses and Position-Specific End Sequencing With Transposon Primers

  • Pradeep K. Chatterjee
  • Joseph C. BakerJr.
Part of the Methods in Molecular Biology™ book series (MIMB, volume 255)


Several protocols for direct sequencing of bacterial artificial chromosome (BAC) and P1 ends have been described recently (1,2). Primers located within a BAC or P1 vector have been used to sequence either end of a genomic insert to facilitate selection of minimally overlapping clones for analyzing large chromosomal regions (1). The entire insert in a BAC or P1-derived artificial chromosome (PAC) clone, however, is usually sequenced using shotgun procedures coupled with gap filling (3). Substantial redundant sequencing of 2-kb subclones appears unavoidable in such approaches. In certain circumstances, however, the sequence of only a small portion of a large BAC or PAC clone is required. Situations such as these arise either when known genetic markers need to be mapped or when new ones need to be identified in a small region of a large clone. Nested deletions become particularly useful in these applications.


Bacterial Artificial Chromosome Luria Bertani Deletion Clone Cotton Gauze Fluorescent Sequencing 
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Copyright information

© Humana Press Inc., Totowa, NJ 2004

Authors and Affiliations

  • Pradeep K. Chatterjee
    • 1
  • Joseph C. BakerJr.
    • 2
  1. 1.Julius L. Chambers Biomedical/Biotechnology Research InstituteNorth Carolina Central UniversityDurham
  2. 2.North Carolina Central UniversityDurham

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