Small-Scale Plasmid DNA Preparation
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For the initial characterization of a recombinant plasmid, it is necessary to determine the size of the plasmid or, preferably, the size and characteristics of the insert itself. A method is therefore required for the simultaneous preparation, from a number of isolates, of plasmid DNA in a state sufficiently pure for restriction enzyme digestion. The requirements of such a procedure are:
| (i) |
A simple method for rapid lysis of the bacterial cells.
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| (ii) |
Separation of plasmid from chromosomal DNA.
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| (iii) |
Removal of proteins and of other components of the cells that might interfere with restriction enzyme treatment.
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| (iv) |
Removal of detergents, salts, etc. used in the process.
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