Sequence-based detection of mutations in cadherin 1 to determine the prevalence of germline mutations in patients with invasive lobular carcinoma of the breast

Table 1 PCR primers and annealing temperature for each of the 16 exons

Coding region	Forward sequence	Reverse sequence	Annealing temperature
Exon 1	ggtgcctccggggctca	gaatgcgtccctcgcaagt	70.5°C
Exon 2	gagtcacccggttccatctacctt	ccagggacaccggcagc	70.5°C
Exon 3	tcttgtctttaatctgtccaatttcctaatctc	gcgcactaaaacaacagcga	63.5°C
Exon 4	tatccgtcttgaattgtcttatcttgttcctcatc	ccctcccagagaaacagagaactt	63.5°C
Exon 5	cagtgttgggatccttctttactaattc	cccggtgtcaacaagcttctaa	63.5°C
Exon 6	tttcttcctcatcagagctcaagt	tccaaagaacctaagagtctttctgagta	69.0°C
Exon 7	tcccaaagtgcagcttgtctaa	taatacacatttgtcctccacaccc	60.5°C
Exon 8	gtcctgacttggttgtgtcg	agacctttctttggaaaccctctaa	69.0°C
Exon 9	agtcttggtactttgtaaatgacacatct	agctgtgaggatgccagtt	69.0°C
Exon 10	aaatgtttcgttttgtttttaacttcattgt	ccagttgctgcaagtcagttga	67.5°C
Exon 11	tttcagctacatgttgtttgctggtcctatt	gctaggaggtcgaggcagcaaag	60.5°C
Exon 12	ttgccaagctgccacattt	gcatggcagttggagcaaag	63.5°C
Exon 13	cctcccctggtctcatcatttc	aagtcaaaggctgagtcacttgc	67.5°C
Exon 14	cttctttatctttggctctcaacactt	agagatcaccactgagctacc	63.5°C
Exon 15	cctactcttcattgtacttcaacct	tgagcttagagatgagccatgc	63.5°C
Exon 16.1	acaggtgtgcccttcctttcactaa	ccaccagcaacgtgatttctgcattt	60.5°C

Cycling conditions were: 95.0°C for two minutes, followed by 50 cycles of 94.0°C for 30 seconds and the specified annealing temperature for 30 seconds, followed by a single cycle of 94.0°C for 30 seconds and then 20.0°C for 30 seconds.

ISSN: 1897-4287