Hi everyone,
Im trying to replicate a specific circular DNA with a single primer, but I only want to amplify the circle once. I know that many polymerases do the rolling circle amplification, but that produces many copies of the circle in the same product.
Is there any polimerase that with one primer amplifies the circle and when the circle is completed stop the replication?
Thanks!
Dear Dr Cabello ,
yours is a very unique situation. I am not sure, but may be Klenow fragment can serve the purpose. although it lacks exonuclease activity (in both directions) and have moderate levels of strand displacement activity too.
regards,
SD
If you just want the sequence at the right length could you use rolling circle amplification then restriction digest with a single site restriction enzyme and you end up with lots of the same length whole circle but in a linear conformation
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