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Abstract Full Text

Article: Branson R and Others. Binge Eating as a Major Phenotype of Melanocortin 4 Receptor Gene Mutations. N Engl J Med 2003;348:1096-1103.

Supplementary Appendix 1

The polymerase-chain-reaction cycling data for melanocortin 4 receptor (MC4R) fragments were as follows: initial denaturation, 2 minutes at 95°C; 7 cycles of 15 seconds at 95°C, 40 seconds at 64°C (0.5 K per cycle), and 90 seconds at 72°C; 36 cycles of 15 seconds at 95°C, 40 seconds at 60°C, and 90 seconds at 72°C; followed by a final extension step of 15 minutes at 72°C. For amplification of the POMC fragments and LEPR sequences, reaction conditions were as follows: initial denaturation, 2 minutes at 94°C; 35 cycles of 30 seconds at 94°C, 30 seconds at 60°C, and 70 seconds at 72°C; followed by a final extension step of 15 minutes at 72°C.