. PCR thermal cycling conditions for recommended protocols. The Pattern column indicates cycling pattern. " S ", for standard reaction pattern, included a starting phase of 2 or 3 minutes at 94 ° C, followed by the cycling phase, with each cycle consisting of 20 - 30 seconds of denaturing at 94 ° C, 20 - 30 seconds of annealing at the annealing temperature, and an extension phase at the temperature specified by the Taq manufacturer. " T ", for touchdown reaction, is like the standard reaction but with three cycling phases, each with a separate annealing temperature. The Cycles column lists the usual number of cycles, with all variant numbers used successfully to generate at least one sequence given in parentheses. For touchdown reactions, the numbers are given for each cycling phase, separated by commas. The Ta column lists the usual annealing temperature in ° C (with less used but successful temperatures in parentheses). For touchdown reactions, the annealing temperatures are given

. PCR thermal cycling conditions for recommended protocols. The Pattern column indicates cycling pattern. “S”, for standard reaction pattern, included a starting phase of 2 or 3 minutes at 94 °C, followed by the cycling phase, with each cycle consisting of 20-30 seconds of denaturing at 94 °C, 20-30 seconds of annealing at the annealing temperature, and an extension phase at the temperature specified by the Taq manufacturer. “T”, for touchdown reaction, is like the standard reaction but with three cycling phases, each with a separate annealing temperature. The Cycles column lists the usual number of cycles, with all variant numbers used successfully to generate at least one sequence given in parentheses. For touchdown reactions, the numbers are given for each cycling phase, separated by commas. The Ta column lists the usual annealing temperature in °C (with less used but successful temperatures in parentheses). For touchdown reactions, the annealing temperatures are given for each cycling phase, separated by commas. The Ext column lists the duration of the extension phase in seconds, with less used but successful durations in parentheses. The Gene column lists the gene(s) successfully amplified with each set of conditions. Subscripts “ex” and “in” refer to conditions used for outer and inner nested (or hemi-nested reactions), respectively.
#PatternCyclesTaExtGeneC1S35 (36)52 (45, 50)7028S, 16S-ND1, COIBC, COIPJC2S3752120wg exC3S35 (37)54150wg inC4T6 (5), 6(5), 36 (35)57, 52, 4590 (60, 120, 180, 300, 330)CAD, TopoC5T6, 6, 3660, 55, 50120 (60, 90, 210, 300)PEPCK exC6S356090 (300)PEPCK in