Construction of fifteen human chromosome-specific DNA libraries from flow-purified chromosomes.
We report the construction of 15 human chromosome-specific DNA libraries. Metaphase chromosomes were purified by flow sorting and the DNA was extracted and cleaved with HindIII before cloning into the lambda vector Charon 21A. A sensitive miniblot hybridization method was used to monitor the physical and biochemical steps in the cloning procedure. Using this method, we have developed a highly efficient protocol for producing large numbers of recombinant phage from 0.2-1.0 X 10(6) sorted chromosomes. DNA from the following chromosomes was cloned: #4, 6, 8, 9, 11, 13, 14 + 15, 16, 17, 18, 19, 20, 21, 22 and Y. These libraries are available to the scientific research community and will be valuable in the genetic analysis of the human genome.
D002460:Cell Line; D002877:Chromosomes, Human / Q000648:ultrastructure*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D005434:Flow Cytometry; D006801:Humans; D007231:Infant, Newborn; D008297:Male; D008677:Metaphase; D009693:Nucleic Acid Hybridization; D012867:Skin / Q000166:cytology
D004247:DNA