The enzyme 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR) is rate-limiting for metabolism of cholesterol; it plays an important role in endogenous cholesterol biosynthesis. We used DNA sequencing technology and created restriction site PCR-RFLP to detect HMGCR SNPs in an F(2) resource population of Gushi chicken and Anka broilers. We found a G/T mutation (Gln/His) in exon 17 and a T/C mutation (Pro/Pro) in exon 18. Based on association analysis of these HMGCR polymorphisms in 864 Gushi/Anka F(2) hybrids, these two mutations have significant effects on growth, carcass, meat quality, and lipid concentration.