Molecular analysis of human chromosome 16 cosmid clones containing NotI sites.
To test the feasibility of using cloned NotI sites as markers for physical mapping, we have screened for cosmid clones spanning the NotI sites on human Chromosome (Chr) 16. Fluorescence in situ hybridization analysis of these clones confirms the previously reported cluster of NotI sites on 16p13.3. Methylation status of the cloned NotI sites on genomic DNA was established by hybridization of the cosmids to Southern blots containing EcoRI and EcoRI/NotI digest of genomic DNA. These results indicated that four of six clones included in our study can be used as linking clones for physical mapping. Two clones have NotI sites which are not cleavable in the cell lines tested. In one clone, the NotI site exists as an isolated rare-cutting restriction enzyme site, whereas in the other clone the NotI site appears to be island-related.
Lerner
T
T
Integrated Genetics, Inc., Framingham, Massachusetts 01701.
Wright
G
G
Leverone
B
B
Dackowski
W
W
Shook
D
D
Anderson
M A
MA
Klinger
K
K
Callen
D
D
Landes
G
G
D015139:Blotting, Southern; D002460:Cell Line; D002885:Chromosomes, Human, Pair 16; D003001:Cloning, Molecular; D003360:Cosmids; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D016521:Electrophoresis, Gel, Pulsed-Field; D015723:Gene Library; D008040:Genetic Linkage; D005819:Genetic Markers; D006801:Humans; D009693:Nucleic Acid Hybridization; D015183:Restriction Mapping
D005819:Genetic Markers; D015252:Deoxyribonucleases, Type II Site-Specific; C038565:GCGGCCGC-specific type II deoxyribonucleases