Rapid mapping of NotI linking clones with differential hybridization and Alu-PCR.
For construction of a NotI restriction map of the human genome, the isolation and mapping of unique NotI linking clones represent important and critical steps. Recently we have shown that an Alu-PCR approach can be used for isolation of NotI linking clones from defined regions of the chromosomes. This represents a useful method for isolating and analyzing a small number of clones, but it would be laborious to use it for mapping many NotI linking clones simultaneously. Here we suggest another modification of Alu-PCR for rapid concurrent mapping of many NotI linking clones. The results clearly demonstrate the utility of this approach. Seventy-one random NotI linking clones were analyzed. Among them, 65 clones (91.5%) were correctly selected and mapped using this approach. With differential hybridization and Alu-PCR, a significant portion of all human NotI linking clones (> 30%) can be rapidly mapped to particular chromosomes or to defined regions of these chromosomes.
Microbiology and Tumor Biology Center, Karolinska Institute, Stockholm, Sweden.
E R
ER
Zabarovsky
V I
VI
Kashuba
I D
ID
Kholodbnyuk
V I
VI
Zabarovska
E J
EJ
Stanbridge
G
G
Klein
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002893:Chromosomes, Human, Pair 3; D017931:DNA Primers; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D008040:Genetic Linkage; D015894:Genome, Human; D006801:Humans; D006822:Hybrid Cells; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid; D015183:Restriction Mapping
D017931:DNA Primers; D015252:Deoxyribonucleases, Type II Site-Specific; C038565:GCGGCCGC-specific type II deoxyribonucleases