Generation of novel sequence tagged sites (STSs) from discrete chromosomal regions using Alu-PCR.
Human DNA segments from discrete chromosomal regions were generated by utilizing Alu-element-based polymerase chain reaction (Alu-PCR) of an irradiation-fusion hybrid containing approximately 10 to 15 Mb of human DNA. Following cloning into a plasmid vector, a subset of the clones was used to generate sequence tagged sites (STSs) de novo. By means of a panel of hybrids containing portions of the human X chromosome, the STSs were shown to localize to two chromosomal regions, Xq24-Xq26 and Xcen-Xq13, reflecting the presence in the irradiation-fusion hybrid of two human chromosome fragments. These results demonstrate that high densities of STSs can be rapidly and efficiently generated from defined regions of the human genome using Alu-PCR.
Cole
C G
CG
Paediatric Research Unit, United Medical School, Guy's Hospital, London, United Kingdom.
Goodfellow
P N
PN
Bobrow
M
M
Bentley
D R
DR
D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D005819:Genetic Markers; D015894:Genome, Human; D006801:Humans; D006822:Hybrid Cells / Q000528:radiation effects; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid; D016324:Sequence Tagged Sites; D014960:X Chromosome
D005819:Genetic Markers