Manipulation of globular DNA molecules for sizing and separation.
Handling large DNA molecules, such as chromosomal DNA, has become necessary due to recent developments in genome science. However, large DNA molecules are fragile and easily broken by shear stress accompanying flow in solution. This fragility causes difficulties in the preparation and handling of large DNA molecules. This study demonstrates the transition of DNA from a coiled to a globular form, which is highly condensed. This state suppresses DNA fragmentation due to shear stress in solution. The transition enables large DNA molecules to undergo mechanical manipulation. We confirmed that the fluorescence intensity of stained globular DNA increases with increasing length, suggesting that the resistance of globular DNA to shear stress is the factor that allows analysis of large DNA by flow cytometry.
Katsura
S
S
Department of Ecological Engineering, Toyohashi University of Technology, Aichi, Japan. katsura@eco.tut.ac.jp
Yamaguchi
A
A
Hirano
K
K
Matsuzawa
Y
Y
Mizuno
A
A
D004271:DNA, Fungal / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification*; D004586:Electrophoresis / Q000379:methods*; D005456:Fluorescent Dyes; D009690:Nucleic Acid Conformation; D013314:Stress, Mechanical
D004271:DNA, Fungal; D005456:Fluorescent Dyes