Table 1 Base sequences of primers used in this study
Forward primer | Reverse primer | |
|---|---|---|
For amplification of 5′ UTR | ||
Canine BRCA2 first PCR | 5′-gggggatccAAAGAAGGTCGGCGGAGGCG-3′ | 5′-gggctcgagCTGCTTGATTGCACCGCGTC-3′ |
Canine BRCA2 nested PCR | 5′-AGAGCGGCACCTCGGAAGG-3′ | 5′-TTTTATCTACAATATTACTCCAGTGCTTGG-3′ |
Human BRCA2 first PCR | 5′-gtcaagcttGTGGCGCGAGCTTCTGAAAC-3′ | 5′-gggctcgagCTGCTTTGTTGCAGCGTGTC-3′ |
Human BRCA2 nested PCR | 5′-GAACTGCACCTCTGGAGCG-3′ | 5′-CCTCCAATGCTTGGTAAATAAGT-3′ |
For quantitative RT-PCR | ||
Canine BRCA2 exon 26–7 | 5′-GACTATGCTTCAGAGCCACACACAG-3′ | 5′-GAAGTCATTTGGGTTGATCCAGGTA-3′ |
Canine BRCA2 exon 1–2 | 5′-GTCAGCTTTCTGGCCGAAGT-3′ | 5′-GTAAATAACTCGCCTTCCGAG-3′ |
Canine RPS18 | 5′-ATAGCCTTTGCCATCACAGCAATTA-3′ | 5′-TTGGTGAGATCGATGTGTCTGCTTTC-3′ |
Human BRCA2 exon 24–5 | 5′-AGGACTTGCCCCTTTCGTCTA-3′ | 5′-TGCAGCAATTAACATATGAGG-3′ |
Human BRCA2 exon 1–2 | 5′-CTCTGGAGCGGACTTATTTACC-3′ | 5′-AATGTTGGCCTCTCTTTGGA-3′ |
Human RPS18 | 5′-TGCGAGTACTCAACACCAAC-3′ | 5′-AGCATATCTTCGGCCCACAC-3′ |
Firefly luciferase | 5′-GATGTACACGTTCGTCACATCTC-3′ | 5′-GACACCTTTAGGCAGACCAGTAG-3′ |
Human BRCA2 exon 27 | 5′-TCATGCCTGTAATCCCAACA-3′ | 5′-AAGATGGGGGTCTCCCTATG-3′ |