A model to describe the size distribution of mammalian genomic fragments recovered by microcloning.
Experiments involving the use of microdissection and microcloning of mammalian chromosomes to obtain genomic clones from individual chromosome regions have demonstrated an aberrant clone recovery. The number average microclone size is well below the empirically observed number average size of genomic restriction fragments. A model is proposed that describes the distribution of microclone sizes by the use of two parameters: (1) the frequency of restriction-enzyme cleavage per bp and (2) the frequency of a second DNA event per bp. The model assumes that any DNA fragment subject to this second event is unclonable. The model shows good fit to the observed microclone size distribution from various experiments when the frequency of the second event is of the order of 0.01. The nature of the second event is unknown but it likely represents a hydrolytic event on the DNA caused by acid fixation of metaphase chromosomes prior to microdissection and microcloning.
Brown
S D
SD
Department of Biochemistry, St. Mary's Hospital Medical School, University of London, Paddington, U.K.
Greenfield
A J
AJ
D000818:Animals; D002875:Chromosomes / Q000032:analysis; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004274:DNA, Recombinant / Q000032:analysis*; D006868:Hydrolysis; D008322:Mammals / Q000235:genetics; D008954:Models, Biological; D008970:Molecular Weight
D004274:DNA, Recombinant; D004262:DNA Restriction Enzymes