Characterization of a cosmid library from flow-sorted chromosomes 11.
A cosmid library specific for human chromosome 11 has been constructed from flow-sorted chromosomes. The flow-purified chromosomes were prepared from the hamster/human hybrid line J1 which contains chromosome 11 as the only human chromosome. Individual clones were sampled in 187 microtitre plates, resulting in a total of 17,952 colonies. Hybridization analysis revealed that 83.7% of these clones were of human and 10.4% of hamster origin. The average insert size was estimated at 33.6 kb, and only 2.4% of insert fragments appear to be rearranged. This should result in 494,487 kb of cloned human DNA representing 3.5 chromosome 11 equivalents. We have prepared high-density nylon membranes of the arrayed library containing 1,536 single colonies per filter. We have demonstrated the usefulness of the library in the molecular genetic analysis of human chromosome 11 by testing for the presence of possibly polymorphic simple repeat motifs, by identifying cosmids that contain inserts from the telomeric ends of chromosome 11 and by assessing the potential of the library for rapid chromosome walking.
Weber
B H
BH
Institut für Humangenetik, Biozentrum, Würzburg, Germany.
Stöhr
H
H
Siedlaczck
I
I
Longmire
J L
JL
Deaven
L L
LL
Duncan
A M
AM
Riess
O
O
D000818:Animals; D002877:Chromosomes, Human; D002880:Chromosomes, Human, Pair 11; D003360:Cosmids; D006224:Cricetinae; D015342:DNA Probes; D005434:Flow Cytometry / Q000379:methods; D015723:Gene Library; D006801:Humans; D006822:Hybrid Cells; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization
D015342:DNA Probes