Therefore, to test the impact of amplicon size on mutation detection, we tested 50 bp (base pair), 100 bp and a 300 bp amplicon on three different mutations (heterozygous p53 mutant I166T (T>C) point mutation, heterozygous RB (retinoblastoma 1) 2 bp deletion, and heterozygous RB 13 bp deletion) designed such that the mutation was in the middle of the amplicon. This evidence concerns the gene RB1 and retinoblastoma.