Genomic DNA from tail biopsies was prepared as described previously 44. Insertion of the pGT0lxf cassette was verified by sequence analysis of PCR-amplified genomic fragments. The following primers were used: A, 5′-GCATCCCTAGCTCTCACTCAGTTAC-3′; and B, 5′-GCGCGTACATCGGGCAAATAA-3′. Genotyping of TORC1 mutant mice was determined by PCR for the wild-type allele and the mutant allele. The wild-type allele was amplified using primers A and C (5′-ATTCCTCATATACCTCTCTTCTGGTGC-3′). The mutant allele was amplified using primers A and D (5′-GCATGAATCAACTTTGGAGACATGCG-3′).
