Abstract:
:A novel method of determining the number of superhelical turns of a covalently-closed plasmid DNA is described. It relies on the determination of the hyperchromicity, and hence the proportion of unstacked basepairs, of a partially heat-denatured sample which co-migrates during electrophoresis with nicked circular duplex DNA. The values obtained for plasmid pBR beta G DNA at 4 degrees C (-29.8 and -33.5 in the two buffers used) agree closely with the values obtained in parallel by topoisomer band-counting. Our method is less precise than band-counting but is readily applicable to determining the superhelicity of very large DNA molecules. Our results confirm earlier findings that magnesium-containing buffers cause an increase in the duplex winding angle, and hence an increase in the number of negative superhelical turns.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Dougherty G,Koller Tdoi
10.1093/nar/10.2.525subject
Has Abstractpub_date
1982-01-22 00:00:00pages
525-38issue
2eissn
0305-1048issn
1362-4962journal_volume
10pub_type
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