Rapid and efficient construction of yeast artificial chromosome contigs in the mouse genome with interspersed repetitive sequence PCR (IRS-PCR): generation of a 5-cM, > 5 megabase contig on mouse chromosome 1.
We have developed a new technique for the generation of YAC contigs in the mouse genome that is based on the ability to detect overlapping clones by hybridization of shared IRS-PCR products. As a demonstration of the technique, a 5-cM, > 5 megabase contig was developed on the distal half of mouse Chromosome (Chr) 1, spanning the region from Lamb2 to At3. The contig covers roughly 5% of the genetic distance of the chromosome and is comprised of more than 80 clones; 71 probes were assigned physical order to the chromosome, of which 59 were new markers generated in this study. Eight of the new probes were shown to be polymorphic between C3H/HeJ-gld and M. spretus. Three probes were mapped on a [(C3H/HeJ-gld x M. spretus) x C3H/HeJ-gld] interspecific backcross to integrate the physical map with a high-resolution genetic map of the region.
Hunter
K W
KW
Center for Cancer Research, Massachusetts Institute of Technology, Cambridge 02139.
Ontiveros
S D
SD
Watson
M L
ML
Stanton
V P
VP
Gutierrez
P
P
Bhat
D
D
Rochelle
J
J
Graw
S
S
Ton
C
C
Schalling
M
M
D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics*; D003001:Cloning, Molecular; D003433:Crosses, Genetic; D017931:DNA Primers / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field; D005260:Female; D008040:Genetic Linkage; D005819:Genetic Markers; D016678:Genome; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D009115:Muridae; D016133:Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid
D017931:DNA Primers; D005819:Genetic Markers