[Construction of a pufferfish gridded cosmid library].
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.
Yang
L
L
Department of Medical Genetics, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 100005.
Liu
S
S
Zhang
X
X
Zhou
W
W
Yang
H
H
D000818:Animals; D003360:Cosmids / Q000235:genetics* / Q000302:isolation & purification; D015698:Genomic Library; D023741:Takifugu / Q000235:genetics*