The position of the gene for the protein actin in the haploid fungus neurospora is known from the complete genome sequence and to verify it one needs to first fulfil the requirements of PCR.
If you had a slow-growing mutant you guessed was an actin mutant and wanted to confirm it, you would be accurate only if the necessary circumstances are satisfied – PCR requires fewer steps. However, for PCR to work, the sequence must be known and the primers must each be present just once in the genome and be sufficiently close together (less than 2 kb). The most effective primers would be those that determine the specificity of the DNA segment amplified.
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