Construction and characterisation of a gridded chicken cosmid library with four-fold genomic coverage.
Gridded genomic libraries are crucial for the positional candidate gene approach. For this purpose we constructed a gridded genomic library from a female chicken using the vector sCos 1. About 110,000 cosmid clones were grown and replicated in 384-well plates. An average insert size of 39 kb was calculated from the analysis of 68 randomly selected clones. No chimerism could be observed from 31 in situ hybridisations. One replica of the library (number 125) has been transferred to the Resource Centre/Primary Database (RZPD) of the German Human Genome Project (DHGP). The whole library was gridded onto four nylon filters at high density for efficient identification of cosmid clones by colony hybridisation. Twenty-two probes were used for screening the library and each of them gave at least one positive signal. This result is in good agreement with a four-fold coverage of the genome as estimated from the insert length and number of recombinant clones. This library provides a powerful tool for rapid physical mapping and complex analysis of the chicken genome.
Buitkamp
J
J
Lehrstuhl für Tierzucht, Technische Universität München, Germany.
Ewald
D
D
Schalkwyk
L
L
Weiher
M
M
Masabanda
J
J
Sazanov
A
A
Lehrach
H
H
Fries
R
R
D000818:Animals; D002645:Chickens / Q000235:genetics*; D002874:Chromosome Mapping / Q000379:methods* / Q000662:veterinary; D003360:Cosmids / Q000737:chemistry*; D004247:DNA / Q000737:chemistry; D005260:Female; D015723:Gene Library; D017404:In Situ Hybridization, Fluorescence / Q000662:veterinary; D008677:Metaphase
D004247:DNA