Table 1 |
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|
RAD51 PCR conditions |
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| Exon |
PCR fragment |
Forward primer |
Reverse primer |
Annealing temperature (°C)a |
Size (bp) |
DHPLC temperature (°C) |
|
|
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| 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 |
|
|
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|
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. |
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|
Lose et al. Breast Cancer Research 2006 8:R26 doi:10.1186/bcr1415 |
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