Changes

PCR genotyping was performed by simultaneous amplification of both wild-type Tsc1 and the deleted allele using the following three primers in a 35 cycle PCR reaction using Perkin Elmer AmpliTaq Gold: F4536, 5′-AGGAGGCCTCTTCTGCTACC-3′; R4830, 5′-CAGCTCCGACCATGAAGTG-3′; and R6548, 5′-TGGGTCCTGACCTATCTCCTA-3′ (Fig. 1D). Products were 295 bp (wild-type) and 368 bp (mutant), and were analyzed on agarose gels.

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