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The price of commercial DNA extraction methods makes the routine use of polymerase chain reaction amplification (PCR) based methods rather costly for scientists in developing countries. A guanidium thiocayante-based DNA extraction method was investigated in this study for the isolation of Escherichia coli (E. coli) DNA from goat, chicken, pig, cow and human stool samples. Two versions of the lysis buffer, with and without α-casein, were tested to alleviate PCR inhibition associated with DNA isolated from stool samples. Results obtained show that, this method using the lysis buffer containing α-casein, produces PCR ready DNA at a fraction of the cost of commercial DNA extraction kits.