A method for rapidly identifying a point mutation in porcine
insulin-like-growth factor 2 intron 7 uses published primers to amplify
the target DNA fragments by polymerase chain reaction. The DNA fragments
are cloned and sequenced for confirmation and method validation. The key
positions of the sequence are modified to generate three primers for
amplifying different DNA fragments with different genotypes by PCR to
avoid additional restriction enzyme digestion. A long primer is used to
specifically amplify the lean muscle mass-enhancing allele, a short
primer is used to specifically amplify the lean muscle mass-suppressing
allele, and the third primer is shared and anneals to the complementary
strand. After PCR and electrophoresis, samples with only the 92 bp band
are identified as the CC genotype, samples with only the 72 bp band are
identified as the GG genotype, and samples with both 92 bp and 72 bp
bands are identified as heterozygotes.