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Updated: Aug 12, 2026

Streamlined Purification of Plasmid DNA From Prokaryotic Cultures
Published on: January 6, 2011
A rapid and complete 4-step protocol for obtaining nucleotide sequence from E. coli genomic DNA from overnight
1Du Pont Merck Pharmaceutical Company, Experimental Station, Wilmington, DE 19880-0328.
Abstract:
We describe a rapid and complete 4-step protocol for obtaining DNA sequence from E. coli genomic DNA starting from overnight cultures. These steps are as follows: isolation and purification of genomic DNA; PCR amplification of the desired region; purification of the amplified DNA product; and finally, direct sequencing using the dideoxy chain termination procedure with simple modifications to improve signal intensity. Clean DNA sequence can be obtained from a large number of strains in a matter of days, using easily available kit-type reagents. This protocol should be generally applicable to genomic DNA from other species.

