Rapid subchromosomal localization of cosmids by nonradioactive in situ hybridization.
A rapid method for localizing large numbers of complete cosmids by nonradioactive in situ hybridization is described. The cosmids are nick translated in the presence of biotin-16-dUTP, incubated with an excess of sonicated human DNA, and used as a probe for in situ hybridization. Sites of hybridization are detected by successive treatments with FITC-labeled avidin and biotinylated anti-avidin antibody. Fifty-two cosmids were localized on chromosome 16 in 5 d relative to translocation breakpoints contained in two cell lines. Rapid identification of chromosome 16 was achieved by cohybridization with a chromosome 16-specific centromeric repeat probe.
Kievits
T
T
Department of Human Genetics, State University of Leiden, The Netherlands.
Dauwerse
J G
JG
Wiegant
J
J
Devilee
P
P
Breuning
M H
MH
Cornelisse
C J
CJ
van Ommen
G J
GJ
Pearson
P L
PL
D002461:Cell Line, Transformed; D002874:Chromosome Mapping; D002885:Chromosomes, Human, Pair 16; D003360:Cosmids; D005260:Female; D006801:Humans; D008856:Microscopy, Fluorescence; D009693:Nucleic Acid Hybridization