Germline mutation analysis of hPMS2 gene in Chinese families with hereditary nonpolyposis colorectal cancer

Table 1 Primer sequences of long-range polymerase chain reaction

Primer name	Sequence (5’-3’)	Size (bp)	Exon
LRPCR1
For	ACGTCGAAAGCAGCCAATGGGAGTT	9964	Exon 1-5
Rev	CTTCCACCTGTGCATACCACAGGCT
LRPCR2
For	GGTCCAGGTCTTACATGCATACTGT	9440	Exon 9
Rev	CTGACTGACATTTAGCTTGTTGACA
LRPCR3
For	GCGTTGATATCAATGTTACTCCAGA	8812	Exon 11, 12
Rev	AGTAGTCAGGGTAAAACATTCCAGT
LRPCR4
For	AAAATTAGTCAGACTTGATGGTGTG	9804	Exon 13-15
Rev	CCTTCCATCTCCAAAACCAGCAAGA

Table 2 Primer sequences and polymerase chain reaction condition of individual exon of hPMS2 gene

Exon	Primer sequence (5’-3’)	Size (bp)	AT (°C)	CN
1	M13F-ACGTCGAAAGCAGCCAATGGGAGTT	475	66	28
	M13R-CAGGTAGAAAGGAAATGCATTCAGT
2	M13F-ACAGTGTTGAGTCATTTCCCACAGT	455	66	28
	M13R-TTCTTAGCATAACACCTGCCTGGCA
3	M13F-TAGTCTGGGCTAGTAAATAGCCAGA	705	68	35
4	M13R-TATGACTTAGATTGGCAGCGAGACA
5	M13F-CTTGATTATCTCAGAGGGATCGTCA	540	68	35
	M13R-TCTCACTGTGTTGCCCAGTCCTAAT
6	M13F-TGCTTCCCTTGATTTGTGCGATGAT	504	67	32
	M13R-TGAGGCAGGAGAATTGCTTGAATCT
7	M13F-ACCCACGAGTTTGACATTGCAGTGA	498	60	35
	M13R-GTAGAGGTTGCAGTGAGCCAAGATA
8	M13F-AGATTTGGAGCACAGATACCCGTGA	414	61	32
	M13R-TGCGGTAGACTTCTGTAAATGCACA
9	M13F-CCTTCTAAGAACATGCTGGTTGGTT	279	64	45
	M13R-ATCTCATTCCAGTCATAGCAGAGCT
10	M13F-AGCCCTTCCGTATTTTGTCTATTCA	719	61	32
	M13R-GCTTTAGAAGCTGTTTGTACACTGT
11	M13F-TCACATAAGCACGTCCTCTCACCAT	1021	64	45
	M13R-GCAACAGAGCAAGACTCTGTCTCAA
12	M13F-GCCAAGATTGTGCCATTGCACTGTA	493	64	25
	M13R-AGTAGATACAAGGTCTTGCTGTGTT
13	M13F-GTGACACTTAGCTGAGTAGTGTTGT	372	64	35
	M13R-ATGTTAGCCAGGCTGGTCTCAAACT
14	M13F-GGTCTGTATCTCCTGACCTCATGAT	473	64	35
	M13R-GCACGTAGCTCTCTGTGTAAAATGA
15	M13F-GCTGAGATCTAGAACCTAGGCTTCT	522	64	35
	M13R-ACACACGAGCGCATGCAAACATAGA

AT: Anneal temperature; CN: Cycle number. The sequence of M13F was 5’-GTAAAACGACGGCCAGT-3’. The sequence of M13R was 5’-AACAGCTATGACCATG-3’.

Table 3 Distribution of single nucleotide polymorphism of hPMS2 gene in 26 probands

Exon	Nucleotide change	Amino acid change	n	SNP (%)
2	c.24-4C>T	-	14	78 (30)
	c.89A>C	Gln30Pro	15
	c.117A>G	Val39Val	15
	c.120G>A	Lys40Ly	4
	c.121G>A	Glu41Lys	15
	c.124T>A	Leu42Ile	8
4	c.288C>T	Ala96Ala	8	18 (6.9)
	c.295A>C	Thr99Pro	10
5	c.406A>G	Met136Val	10	60 (23.1)
	c.418A>G	Asn140Asp	10
	c.429T>C	Ile143Ile	10
	c.452G>A	Arg151His	10
	c.478C>A	Gln160Lys	10
	c.492C>T	Ser164Ser	10
11	c.1408C>T	Pro470Ser	7	24 (9.2)
	c.1454C>A	Thr485Lys	11
	c.2006+6G>A	-	7
12	c.2007-4G>A	-	11	12 (4.6)
	c.2007-7C>T	-	1
13	c.2253T>C	Phe751Phe	1	1 (0.4)
14	c.2324A>G	Asn775Ser	3	5 (1.9)
	c.2340C>T	Pro780Pro	2
15	c.2466T>C	Leu822Leu	12	36 (13.8)
	c.2570G>C	Gly857Ala	2
	c.92dupA	-	17
	c.17G>C	-	5

SNP: Single nucleotide polymorphism.