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large plasmid isolation

large plasmid isolation - Basic Lab Protocols and Techniques

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Old 02-01-2010, 06:14 AM
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Thumbs up large plasmid isolation



Hi ,iam isolating plasmid of 220 Kb size by both kit and manual method(from single colony).After isolation iam frequently getting 5-6 bands!!Wat is the reason?is there any other protocol?
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Old 02-03-2010, 05:10 PM
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Default Re: large plasmid isolation

Do you have a picture of your gel? Plasmids can show up as different sized bands depending on the amount of super-coiling. Cut your plasmid with an enzyme with a single site in your vector. You should get 1 band. If not, then you have problems...
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Old 02-08-2010, 10:42 AM
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Default Re: large plasmid isolation

Hi thanks for ur reply.Im isolating 220 kb virulence plasmid of shigella flexneri.I donno y after isolation by both kit and manual method ,each time getting 5-7 bands.i wil send you plasmid photo.
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