High molecular weight DNA from nucleated erythrocytes for use in pulsed-field gel electrophoresis (PFGE).
Nucleated erythrocytes of lower vertebrates provide a source of genomic DNA that can be used in pulsed-field gel electrophoresis (PFGE) studies. Difficulties reported in the preparation of chicken erythrocyte DNA for analysis by PFGE suggest that the presence of hemoglobin iron may result in iron-mediated DNA degradation. We report here modifications to the procedures established for isolation of high molecular weight DNA from mammalian cells. By increasing the volume of buffers and extending the incubation periods to allow for the removal of hemoglobin iron, we have successfully prepared channel catfish erythrocyte DNA suitable for analysis by PFGE.
Ventura-Holman
T
T
Department of Microbiology, University of Mississippi Medical Center, Jackson 39216, USA.
Lobb
C J
CJ
D000818:Animals; D004247:DNA / Q000302:isolation & purification*; D016521:Electrophoresis, Gel, Pulsed-Field; D004912:Erythrocytes / Q000737:chemistry*; D007059:Ictaluridae; D008970:Molecular Weight
D004247:DNA