Direct microdissection and microcloning of a translocation breakpoint region, t(1;11)(q42.2;q21), associated with schizophrenia.
We describe the generation of large-fragment microclone libraries from the chromosomal breakpoint of a reciprocal balanced translocation linked to schizophrenia. The abnormality was visible under the phase-contrast microscope, allowing direct dissection from unstained, unbanded metaphases. Two separate microdissection experiments yielded 443 and 672 recombinants, respectively. Following complete EcoRI digestion, inserts with an average size of 0.3 kb (range, 0.2-3 kb) were obtained in the first experiment and 1.5 kb (range, 0.15-6.5 kb) in the second. FISH analysis of pooled clones "painted" back onto the derivative chromosome and assignment of microclones to somatic cell hybrids confirmed the fidelity of the method. Microdissection of chromosome regions identified by karyotype rearrangements in unstained, unbanded metaphases is a potentially powerful tool for positional cloning.
Muir
W J
WJ
MRC Human Genetics Unit, Western General Hospital, Edinburgh, Scotland, UK.
Gosden
C M
CM
Brookes
A J
AJ
Fantes
J
J
Evans
K L
KL
Maguire
S M
SM
Stevenson
B
B
Boyle
S
S
Blackwood
D H
DH
St Clair
D M
DM
D001483:Base Sequence; D002874:Chromosome Mapping; D002878:Chromosomes, Human, Pair 1; D002880:Chromosomes, Human, Pair 11; D003001:Cloning, Molecular / Q000379:methods*; D004210:Dissection / Q000379:methods*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D008846:Micromanipulation; D008969:Molecular Sequence Data; D012559:Schizophrenia / Q000235:genetics*; D014178:Translocation, Genetic