Fluorescence in situ hybridization mapping of human chromosome 19: mapping and verification of cosmid contigs formed by random restriction enzyme fingerprinting.
Automated restriction enzyme fingerprinting of 7900 cosmids from chromosome 19 and calculation of the likelihood of their overlap based on shared fragments have resulted in the assembly of 743 sets of overlapping cosmids (contigs). We have mapped 22% of the formed contigs (n = 165) and all of the contigs with minimal tiling paths exceeding 6 members (n = 50) to chromosomal bands by fluorescence in situ hybridization using DNA from at least one member cosmid. The estimated average size of the formed contigs is 60-70 kb. Thus, members of a correctly formed contig are expected to lie close to each other in metaphase and interphase chromatin. Therefore, we tested the contig assembly process by comparing the band assignment of two or more members selected from each of 97 contigs. Forty-two of these contigs were further characterized for valid assembly by determining the proximity of members in interphase chromatin. Using these tests, we surveyed a total of 431 joins counted along the minimal tiling path (280 in interphase as well as metaphase) and found 6 erroneous joins, one in each of 6 contigs (6% of tested).
Trask
B
B
Human Genome Center, Lawrence Livermore National Laboratory, Livermore, California 94550.
Christensen
M
M
Fertitta
A
A
Bergmann
A
A
Ashworth
L
L
Branscomb
E
E
Carrano
A
A
Van Den Engh
G
G
D002888:Chromosomes, Human, Pair 19; D003360:Cosmids / Q000235:genetics*; D016172:DNA Fingerprinting; D004262:DNA Restriction Enzymes; D005453:Fluorescence; D015723:Gene Library; D006801:Humans; D017403:In Situ Hybridization
D004262:DNA Restriction Enzymes