Table 1. Primers used in this study for cDNA clone library construction and real-time PCR quantification of arsenic biotransformation-related genes (arrA and arsB/acr3) and the housekeeping gene recA.
Target | Primer set | Forward primer (5'–3') | Reverse primer (5'–3') | Annealing temperature (°C) | Amplicon size (bp) | Reference or source | |
---|---|---|---|---|---|---|---|
arrA | Clone library | HAArrA-D1F/HAArr-G2R | CCGCTACTACACCGAGGGCWWYTGGGRNTA | CGTGCGGTCCTTGAGCTCNWDRTTCCACC | 53.5 | ∼500 | Kulp et al., 2006 |
Real-time PCR | qArr1F/qARR1R | GATCCACGCTTCTCCACCTC | CCCCGGCTTTAAAGAGGTTC | 60 | ∼172 | This study | |
arsB | Clone library | AarsB1F/AarsB1R | GAACATCGTCTGGAAYGCNAC | GTACACCACCAGRTACATNCC | 55 | ∼750 | Achour et al., 2007a |
acr3-1 | Clone library | Aacr1F/Aacr2R | GGCCTGATCGTNATGATGTAYCC | GCGATGGCCAGCTCRAARTTRTT | 57b | ∼750 | Achour et al., 2007a |
Real-time PCR | qA1-3F/qA1-2R | ATGGCCAGCTCGAAaTTGTT | 60 | ∼94 | This study | ||
acr3-2 | Clone library | dacr5F/dacr4R | TGATCTGGGTCATGATCTTCCCVATGMTGVT | CGGCCACGGCCAGYTCRAARAARTT | 52 | ∼750 | Achour et al., 2007a |
Real-time PCR | qA2-1F/qA2-1R | GGCCAGTTCGAAGAAGTTGG | GCCGATTTTGATCCAGGTGT | 60 | ∼127 | This study | |
recA | Clone library | recA-48F/recA-583R | CCAGATHGARAARCAGTT | TTCATACGGATCTGGTTGAT | 55 | ∼540 | This study |
Real-time PCR | qrecA-2F/qrecA-3R | TTGAGAAGCAGTTCGGCAAA | CGGTCCGAAGATCTCGATG | 60 | ∼156 | This study |
Primers modified by Fahy et al., unpublished.
57°–0.5° for 10 cycles, 52° for 30 cycles.