Whole length analysis of the c-KIT gene
The coding region of c-KIT was sequenced, as reported by Fontanesi et al(Fontanesi et al., 2010). The sequence was aligned against the pig genomic sequence AC141857 (clone RP44-473N18) containing 21 c-KIT exons using BLASTN analysis (http://blast.ncbi.nlm.nih.gov/Blast.cgi). The splice mutation site in exon 17 was amplified with primer KIT17, designed by Marklund et al(Marklund et al., 1998). The 4-bp indel in intron 18 was amplified using primer Intron 18, and the 164/168 bp product was sequenced. The thermal cycling profile was 95 °C for 2 min, followed by 35 cycles of 95 °C for 10 s, 58 °C for 10 s, and 72 °C for 10 s, with a 5 min extension at 72 °C and holding at 4 °C. PCR products were purified by 2% gel electrophoresis and sequenced.