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Table 3 Effects of PCR-related factors on genotyping errors of HER2I655V AS-PCR for AG genotype

From: Modified allele-specific PCR improves HER2 Ile655Val detection by reducing genotyping errors

Type of error   DNA template ratio (w/w)a Annealing temperature (°C)b
   A:G A:G A:G A:G A:G 51.1 57.7
   1:1 1:0.25 0.25:1 0.5:0.5 0.25:0.25
Experiments   1th 2nd 1th 2nd 1th 2nd 1th 2nd 1th 2nd 1th 2nd 1th 2nd
LDO No PCR product 0 0 0 0 1 0 0 0 2 1 1 2 0 0
ADO AA genotype only 0 0 1 1 0 0 0 0 0 0 2 0 0 1
GG genotype only 0 1 0 0 1 2 0 0 0 0 0 0 0 0
Correct genotype 5 4 4 4 3 3 5 5 3 4 2 3 5 4
Total 5 5 5 5 5 5 5 5 5 5 5 5 5 5
Mean of genotyping error rate (%) 10% 20% 40% 0% 30% 50% 10%
  1. Allele-specific PCR was done with 5 replicates in two independent experiments
  2. aAllele-specific PCR was done using annealing temperatue of 54.3 °C
  3. bAllele-specific PCR was done using mixed DNA template of 1 ng AA genotype and 1 ng GG genotype