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PubMed · 1954014

A rapid and complete 4-step protocol for obtaining nucleotide sequence from E. coli genomic DNA from overnight cultures.

S Seetharam·I B Dicker
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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.

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Base SequenceDNA, BacterialEscherichia coliPolymerase Chain ReactionTime FactorsJournal Article

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S Seetharam, I B Dicker. 1991. A rapid and complete 4-step protocol for obtaining nucleotide sequence from E. coli genomic DNA from overnight cultures.. https://pubmed.ncbi.nlm.nih.gov/1954014/

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