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[Construction of a pufferfish gridded cosmid library]
1Department of Medical Genetics, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 100005.
Objective:
Pufferfish is a newly-established model organism in human genome research, which has been demonstrated its unique advantages in comparative genomics. Construction of a genomic library of pufferfish is the prerequisite to any studies of its genome.
Methods:
Genomic DNA from Fugu rubripes, a species distribute only in Japan and China seas, was used to construct a library in a novel cosmid vector, named sCOGH2, which contains an exon-trapping cassette.
Results:
This library is composed of 57,600 clones gridded in 60,096-well cell culture plates, one clone each well. The average size of the inserts is around 35 kb. It provides 99.2% probability to find any unique DNA fragment of pufferfish in this library. The clones of this library can survive after more than 10 times freeze-thaw, also demonstrating high stability in propagation. The clones shone highly positive signals when hybridized with the pufferfish genomic DNA.
Conclusions:
The library accords with qualitative demands of a cosmid library.