Construction of a radiation hybrid map of chromosome 9p.
A radiation hybrid panel has been constructed for chromosome 9 using the somatic cell hybrid GM10611 as the donor cell line fused to the hamster cell line A23. The hybrid GM10611 was characterized by fluorescence in situ hybridization and reverse painting onto spreads of normal human metaphase chromosomes; it contains human chromosome 9 as the only cytogenetically detectable human material. GM10611 was irradiated with 6000 rads of X rays prior to fusion, a total of 93 independent clones were selected, and frozen stocks and DNA were prepared from each clone. These clones were screened by PCR amplification with oligonucleotide primers for sequence-tagged sites specific for 50 single-copy loci mapping to the short arm of chromosome 9. The average retention frequency of these hybrids was approximately 23%. The markers were ordered into a framework map by analyzing coretention patterns, minimizing the number of obligatory chromosome breaks, and finally confirming the order by maximum likelihood methods. A framework map ordering 27 markers with odds greater than 1000:1 was constructed. A further 16 markers that could not be uniquely placed on the map with the required support were positioned within a range of adjacent intervals.
Bouzyk
M
M
Human Genetic Resources, Clare Hall Laboratories, South Mimms, Hertsfordshire, EN6 3LD, United Kingdom.
Bryant
S P
SP
Schmitt
K
K
Goodfellow
P N
PN
Ekong
R
R
Spurr
N K
NK
D000818:Animals; D001483:Base Sequence; D002459:Cell Fusion; D002460:Cell Line; D002874:Chromosome Mapping; D002899:Chromosomes, Human, Pair 9; D006224:Cricetinae; D017931:DNA Primers; D005819:Genetic Markers; D006801:Humans; D006822:Hybrid Cells / Q000528:radiation effects; D017404:In Situ Hybridization, Fluorescence; D007621:Karyotyping; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D014965:X-Rays
D017931:DNA Primers; D005819:Genetic Markers