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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)

Abstract

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.

Keywords

Bacterial Artificial Chromosome Luria Bertani Deletion Clone Cotton Gauze Fluorescent Sequencing 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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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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