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[Construction of a pufferfish bacterial artificial chromosome library]
Objective:
Study of human genome by taking advantage of investigating pufferfish's (Fugu rubripes), compact model vertebrate genome.
Methods:
Construct a pufferfish bacterial artificial chromosome (BAC) library using Japanese pufferfish (Fugu rubripes) sperm DNA and pBelo BAC11 vector.
Results:
The library consists of 23,040 clones, which have been arrayed in 240,96-well microtiter plates. 100 BAC clones have been selected at random and analyzed for estimation of the average insert size, which turns out to be 100 kb indicating its nearly 6-fold coverage of pufferfish haploid genome. Restrict analysis of 10 BAC clones show that these inserts are stable even propagated for at least 100 cell generations.
Conclusions:
The library accords with the demands of a BAC library.