High molecular weight DNA from fixed cytogenetic preparations.
Cell pellets that have been stored in routine clinical cytogenetic fixative were studied for the presence of intact DNA. A method for the isolation of high molecular weight DNA from fixed cytogenetic preparations of human leukocytes, bone marrow, and cell hybrid cultures is presented. DNA preparations from fixed pellets were cleaved with restriction enzymes, transferred to nitrocellulose filters after agarose gel electrophoresis, and hybridized to radiolabeled probes to demonstrate that fixed cell pellets could yield DNA of sufficient quality for Southern blot hybridization analysis. This protocol may be useful for molecular analysis of DNA from fixed cell pellets of patients who are unavailable for additional sampling.
D001853:Bone Marrow / Q000032:analysis; D003581:Cytodiagnosis / Q000379:methods*; D004247:DNA / Q000302:isolation & purification*; D005404:Fixatives; D005615:Freezing; D006801:Humans; D006822:Hybrid Cells / Q000032:analysis; D007962:Leukocytes / Q000032:analysis; D008970:Molecular Weight; D013048:Specimen Handling
D005404:Fixatives; D004247:DNA