How to detect the copy number of plasmid in bacteria?!

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wei
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How to detect the copy number of plasmid in bacteria?!

hello,
I am searching the answer for title. Could someone give any approach to detect this. I so appreciate. Thanks a lot.. :)

Tony Rook
Tony Rook's picture
Wei:

Wei:

The most efficient way to determine bacterial copy number is by using real time PCR. If you do not have access to a real time PCR thermocycler you can also estimate copy number using a gel quantification assay (Pushnova 2000).

Here are some good references for quantitative real time PCR methods:

Lee CL, et al. Quantitative real-time polymerase chain reaction for determination of plasmid copy number in bacteria. Journal of Microbiological Methods 65 (2006) 258 267.

Lee C, et al. Absolute and relative QPCR quantification of plasmid copy number in Escherichia coli. Journal of Biotechnology 123 (2006) 273280.
http://www.aseanbiotechnology.info/Abstract/21020445.pdf

Here are some good reference for estimation of copy number via gel quantification:

Pushnova, EA, et al. An Easy and Accurate Agarose Gel Assay for Quantitation of Bacterial Plasmid Copy Numbers. Analytical Biochemistry 284, 7076 (2000).

Schmidt T, et al. Rapid Determination of Plasmid Copy Number. J of Biotechnology, 1996 49: 219-229.

Please update the board on the outcomes of your experiments.

Good Luck!