Gel Origin
Gel Origin
Gel Origin
OC DNA
CCC Supercoiled DNA
CCC Relaxed DNA
OC DNA + CCC Relaxed DNA
Linear DNA
CCC Supercoiled DNA
OC DNA Topo Relaxation Products: Topoisomers (relaxed DNA forms that differ in linking number)
CCC Supercoiled DNA
Panel A: Typical EB containing gel separation of the 3 DNA forms of Plasmid DNA. Note that the resolution between supercoiled and relaxed MAY BE very poor if the gel is run fast for short times. In general you can resolve these forms but it takes longer run times. ALWAYS include markers (known supercoiled and known relaxed DNA species).
Panel B: Typical Non-EB containing gel showing the separation of the different plasmid DNA forms. Resolution of supercoiled DNA from nicked (open cirucular, OC) and relaxed (covalently closed circular, ccc) DNA is excellent. NOTE: Relaxed CCC DNA and nicked OC DNA may resolve poorly (if at all) in this system. These types of gels are ideal if you want to quickly test for relaxation activity. They are less than ideal for detecting OC nicked DNA products that arise from topo I action on DNA in the presence of drugs or poisons like camptothecin.
Panel C: Typical Non-EB containing gel separation of OC Nicked DNA, ccc Supercoiled DNA and and a gaussian distribution of topoisomers. The formation of these topoisomerase are diagnostic for topo I or II activity. For this reason, these gels are ideal for activity measurements.
Dedicated to studies of topoisomerase enzymes.
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