Specific Cloning of DNA Fragments Absent from the DNA of a Male Patient with an X Chromosome Deletion

@article{AbsentFragments, abstract = {A method that allows the specific cloning of DNA fragments absent from patients homozygous or hemizygous for chromosomal deletions is described. The method involves phenol-accelerated competitive DNA reassociation and subsequent molecular cloning of appropriately reassociated molecules. The deletion DNA sample utilized in the competition was isolated from a patient with a minute interstitial deletion in the short arm of the X chromosome. Sheared DNA isolated from a male child, who was diagnosed as having Duchenne muscular dystrophy, chronic granulomatous disease, and retinitis pigmentosa, was combined in a 200-fold excess with Mbo I-cleaved DNA isolated from a 49,XXXXY human lymphoid cell line, and the mixture was subjected to a phenol-enhanced reassociation technique. Analysis of 81 unique segments derived from cloned reassociated DNA molecules has led to the identification of 4 (5\%) human DNA fragments that are absent from the male patient's DNA. The 4 clones were localized, on the basis of hybridization with restriction nuclease-digested genomic DNA from a panel of human and human-rodent hybrid cell lines, into three regions surrounding band 21 of the short arm of the normal human X chromosome. These clones are potential linkage markers for the diseases affecting this boy. Each clone, as well as others obtainable by this approach, may also serve as a starting point in the eventual cloning of these three X-linked-disease loci. Extension of this approach to other loci, including human tumors potentially homozygous for small deletions, should also be possible.}, author = {Kunkel, Louis M. and Monaco, Anthony P. and Middlesworth, William and Ochs, Hans D. and Latt, Samuel A.}, citeulike-article-id = {1124395}, doi = {10.1073/pnas.82.14.4778}, journal = {PNAS}, keywords = {absent, dna, nullomers}, month = {July}, number = {14}, pages = {4778--4782}, posted-at = {2007-02-27 01:12:51}, priority = {3}, title = {Specific Cloning of DNA Fragments Absent from the DNA of a Male Patient with an X Chromosome Deletion}, url = {http://dx.doi.org/10.1073/pnas.82.14.4778}, volume = {82}, year = {1985} }

TY - JOUR ID - AbsentFragments L3 - citeulike-article-id:1124395 N2 - A method that allows the specific cloning of DNA fragments absent from patients homozygous or hemizygous for chromosomal deletions is described. The method involves phenol-accelerated competitive DNA reassociation and subsequent molecular cloning of appropriately reassociated molecules. The deletion DNA sample utilized in the competition was isolated from a patient with a minute interstitial deletion in the short arm of the X chromosome. Sheared DNA isolated from a male child, who was diagnosed as having Duchenne muscular dystrophy, chronic granulomatous disease, and retinitis pigmentosa, was combined in a 200-fold excess with Mbo I-cleaved DNA isolated from a 49,XXXXY human lymphoid cell line, and the mixture was subjected to a phenol-enhanced reassociation technique. Analysis of 81 unique segments derived from cloned reassociated DNA molecules has led to the identification of 4 (5%) human DNA fragments that are absent from the male patient's DNA. The 4 clones were localized, on the basis of hybridization with restriction nuclease-digested genomic DNA from a panel of human and human-rodent hybrid cell lines, into three regions surrounding band 21 of the short arm of the normal human X chromosome. These clones are potential linkage markers for the diseases affecting this boy. Each clone, as well as others obtainable by this approach, may also serve as a starting point in the eventual cloning of these three X-linked-disease loci. Extension of this approach to other loci, including human tumors potentially homozygous for small deletions, should also be possible. IS - 14 JF - PNAS EP - 4782 TI - Specific Cloning of DNA Fragments Absent from the DNA of a Male Patient with an X Chromosome Deletion VL - 82 SP - 4778 KW - absent KW - dna KW - nullomers AU - Kunkel, Louis AU - Monaco, Anthony AU - Middlesworth, William AU - Ochs, Hans AU - Latt, Samuel PY - 1985/07/15/ UR - http://dx.doi.org/10.1073/pnas.82.14.4778 ER -