Polymerase chain reaction-single strand conformational polymorphism analysis of rearranged during transfection proto-oncogene in Chinese familial hirschsprung’s disease

Table 1 Primers used in PCR amplification and DNA sequencing experiments.

Exon	Sequence (5'→3')	Annealing t (°C)	Product size (bp)
11	F: ACACCACCCCCACCCACAGAT	62	273
	R: AAGCTTGAAGGCATCCACGG
13	F: GACCTGGTATGGTCATGGA	58	253
	R: AAGAGGGAGAACAGGGCTGTA
15	F: GACTCGTGCTATTTTTCCTAC	60	234
	R: TATCTTTCCTAGGCTTCCC
17	F: CCCCACTAGATGTATAAGGG	59	232
	R: TCACTGGTCCTTTCACTCTCT

F: forward primer; R: reverse primer.

Table 2 Mutated sites observed in RET proto-oncogenes in 48 sporadic HD patients.

Case	Sex	Range of aganglionic segment	Exon	Nucleotide change	Amino acid change	Mutation types
1	M	Long-segment	11	G15165→A	G667S	Missense mutation
2	M	Short-segment	11	G15165→A	G667S	Missense mutation
3	F	Long-segment	11	G15165→A	G667S	Missense mutation
4	M	Long-segment	13	T18888→G	L745L	Silent mutation
5	M	Long-segment	13	A18919→G	K756E	Missense mutation
6	M	Long-segment	13	18974insG	- - - -	Frameshift mutation
7	M	Short-segment	13	18974insG	- - - -	Frameshift mutation
8	M	Short-segment	15	G20692→A	Q916Q	Silent mutation

M: male; F: female.