A shortened PCR procedure was developed in a conventional thermal cycler. Overnight cultures of E. coli were used for PCR to amplify fragments of the uidA and slxII genes. A standard PCR program (30 cycles of 1 m denaturation at 94C, 1 m annealing at 54C or 60C, and 2.5 m elongation at 72C) required 3.5 h to complete and was able to detect a product from 1 × 103 PCR template cells in an agarose gel. The shortened PCR program (35 cycles of 17 s denaturation at 94C, 20 s annealing at 54C or 60C and 23 s elongation at 72C) required 1.5 h time and was able to detect PCR product from 5×10 PCR template cells. The results show that it is possible to shorten the time of each PCR cycle producing a more rapid method for the detection of E. coli while still using a conventional thermal cycler. This approach was successful using different primer pairs, indicating other researchers could use this approach to significantly shorten their PCR reaction times.