Detection of germline mutations of hMLH1 and hMSH2 based on cDNA sequencing in China

Table 1 Sequence and localization of primers used for amplification of cDNA of hMLH1 and hMSH2

Sense	Antisense
hMLH1-1F(1-18)	hMLH1-5R(2198-2175)
CTTGGCTCTTCTGGCGCC	GAGCGCAAGGCTTTATAGACAATG
hMLH1-4F(1333-1353)	hMLH1-6R(2484-2459)
GCTGAAGTGGCTGCCAAAAAT	TATGTTAAGACACATCTATTTATTTA
hMSH2-1F(1-21)	hMSH2-7R(2753-2732)
GGCGGGAAACAGCTTAGTGGG	GGGCATTTGTTTCACCTTGGAC
hMSH2-6F(1898-1920)	hMSH2-8R(3145-3114)
CGTGTCAAATGGAGCACCTGTTC	CCACAAACTACATGATTTTATTTATAAAATTC

Table 2 hMLH1 and hMSH2 primers used for sequencing of cDNA

Sense	Antisense
hMLH1-1F CCTGGCTCTTCTGGCGCC	hMLH1-1R CTTTTCTCCTCGTGGCTATGTTGT
hMLH1-2F ATGTGCTGGCAATCAAGGGA	hMLH1-2R GGTGCACATTAACATCCACATTCT
hMLH1-3F CCAAAAACACACACCCATTCCT	hMLH1-3R CCTTTGTTGTATCCCCCTCCA
hMLH1-4F GCTGAAGTGGCTGCCAAAAAT	hMLH1-4R CATCTTCCTCTGTCCAGCCACTC
hMLH1-5F TTGCCATGCTTGCCTTAGATAGTC	hMLH1-5R GAGCGCAAGGCTTTATAGACAATG
hMLH1-6F GCTCCATTCCAAACTCCT	hMLH1-6R TATGTTAAGACACATCTATTTATTTA
hMSH2-1F GGCGGGAAACAGCTTAGTGGG	hMSH2-1R CTCTGGCCATCAACTGCGGAC
hMSH2-2F GGCTTCTCCTGGCAATCTCTCTCA	hMSH2-2R CTTGATTACCGCAGACAGTGATGAAAC
hMSH2-3F GCAAAAAGGGAGAGCAGATGAATAGTG	hMSH2-3R GGCAAGTCGGTTAAGATCTGGGAAT
hMSH2-4F AGATGCAGAATTGAGGCAGACTTTACA	hMSH2-4R GGACTTTTTCTTCCTTACAGGTTACACG
hMSH2-5F CAGAGATCTTGGCTTGGACCCT	hMSH2-5R TTCAACACAAGCATGCCTGGAT
hMSH2-6F CGTGTCAAATGGAGCACCTGTTC	hMSH2-6R GATTGGCCAAGGCAGTAAGTTCAT
hMSH2-7F AATCATAGATGAATTGGGAAGAGGAACT	hMSH2-7R GGGCATTTGTTTCACCTTGGAC
hMSH2-8F CTATCTGGAAAGAGAGCAAGGTGAA	hMSH2-8R CCACAAACTACATGATTTTATTTATAAAATTC

Table 3 hMLH1 and hMSH2 mutations detected by cDNA sequencing

Families	Genes	Exon	Codons affected	DNA change	Amino acid change	Mutation types
H31	hMLH1	16	581	T>C, at 1742	Pro→Leu	Missense
H111	hMLH1	19	Non-coding area	A>T, at 2438
H114	hMLH1	12	384	T→A, at 1151	Val→Asp	Missense
H2	hMLH1	8	217	T->C, at 649	Arg→Cyc	Missense
H11	hMLH1	1	5	A>C, at 14	Pro→His	Missense
H38	hMLH1	2	99	A>C, at 295	Arg→Arg	Silent
H38	hMLH1	2	99	Del G, at 296	Frame shift	Frame shift