Table 1

RAD51 PCR conditions

Exon
PCR fragment
Forward primer
Reverse primer
Annealing temperature (°C)a
Size (bp)
DHPLC temperature (°C)

1
1
GCAAGCGAGTAGAGAAGTGGA
AACTGCCGCTGAGCACTG
54
218
65
2
2
ATGGCCTTGGCTTTTCCTAA
GGCCCTGCCAGACATATTTA
54
391
57
3
3
TGGAACCAACTTCCCATCTC
TTCCCACTAATGCCTCCCTA
54
341
58
4
4
CTCTTCCCATTGCACACCTT
CACCTGGCCTTCCTCTATCTC
54
371
52, 57
5
5
TCTGATGAGCTCCAAGAACA
TGACATGGAAGGATTTTGAAG
51
344
58
6
6
AAGGGAATGCCTCCTTCCTA
CCAAACTAACCCTGGCAATC
54
314
59
7
7
CAAATTGCTCATCTGCCTG
TGAGGCACCGTTTAACAAGA
54
397
59
8
8
TGGTAAGGAAGGGACCAGAA
TGTGGCCATAGACACTCCAA
57
390
60
9
9
TCGTTATTTTGTGGGGGAAA
ACAGGGGAGAGGCATATCAA
54
447
58
10
10a
TTGGTGCTTTGGTCTGTGTC
ATACCCCTCCTCCAAAACCA
54
409
59
10
10b
CAGGAGACAGGTCAGTAGTCACA
AGGTTTGGCACAAGACTCCA
51
374
57
10
10c
TGATCTTGTGTAAGGGTTTGGTT
GCAATCTCGACTCACTGCAA
51
375
60
10
10d
GATAGCCTGAGGTGGGAGAA
TCTGCAAGTGGGACTTTCCT
54
319
60

a'Touchdown' PCR amplification conditions were as follows: denaturation at 94°C for 10 minutes, followed by two cycles of 94°C for 30 seconds, 30 seconds at the fragment annealing temperature (TA) + 6°, and 72°C for 30 seconds. The conditions remained the same for the rest of the PCR except for the annealing temperature, which consisted of two cycles at (TA + 4°C), then two cycles at (TA + 2°C) and, finally, 35 cycles at the TA. A final extension step was conducted at 72°C for 7 minutes.

Lose et al. Breast Cancer Research 2006 8:R26   doi:10.1186/bcr1415

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