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- W197152009 abstract "Recently, we have developed new procedures for the cloning of large DNA fragments using a bacteriophage P1-derived vector, pCYPAC1. In view of the large sizes (up to 400 kb clones have been observed) and the single copy mode of maintenance, we have designated our clones as {open_quotes}PACs{close_quotes} for {open_quotes}P1-derived artificial chromosomes{close_quotes}. Our subsequent efforts have focused on the construction of a large human PAC library from MboI digested human DNA (from white blood cells). About 120,000 PAC clones have been picked into 312 microtiter dishes of 384 wells. Preliminary characterization has revealed that the insert containing clones have average inserts of 110-120 kb and that the library has approximately 27% empty clones (vector without insert). Based on the average insert size and the total number of recombinant clones, we estimate that the library has 3-fold genome redundancy. This estimate is consistent with results obtained by screening of the library: 18 STS markers detect 52 corresponding clones and 6 unique probes find 20 positives. The library is expanded to a final size of 6- to 8-fold genome redundancy and is also being characterized with by fluorescent in situ hybridization and chromosome-walking procedures. Copies of the arrayed library are being distributed." @default.
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- W197152009 date "1994-09-01" @default.
- W197152009 modified "2023-09-23" @default.
- W197152009 title "Preparation and characterization of a 3-fold redundant human PAC library" @default.
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