Table 1 Primers used for 3′-, 5′-RACE, ORF cloning, and plasmid construction
From: Gene structures, biochemical characterization and distribution of rat melatonin receptors
Gene | Direction | Primer sequences |
|---|---|---|
3′-RACE | ||
Universal | ||
RT primer | Reverse | 5′-GCTGTCAACGATACGCTACGTAACGGCATGACAGTG(T)18-3′ |
1st PCR | Reverse | 5′-GCTGTCAACGATACGCTACGTAACG-3′ |
2nd PCR | Reverse | 5′-CGCTACGTAACGGCATGACAGTG-3′ |
MT1 | ||
1st PCR | Forward | 5′-AAACCGGACAGCAAACCCAAACT-3′ |
2nd PCR | Forward | 5′-GGGCCCCACTCAACTTCATAGGTC-3′ |
MT2 | ||
1st PCR | Forward | 5′-CCGAAGGAAGGCAAAGGCTGAGAG-3′ |
2nd PCR | Forward | 5′-CCAGAAGGGCTTTTTGTCACCAG-3′ |
5′-RACE | ||
MT1 | ||
RT primer | Reverse | 5′-pAGTAACTAGCCACGAAGA-3′ |
1st PCR | Forward | 5′-AAACCGGACAGCAAACCCAAACT-3′ |
Reverse | 5′-GCCCAGGATGTCCACCACGATAGT-3′ | |
2nd PCR | Forward | 5′-GGGCCCCACTCAACTTCATAGGTC-3′ |
Reverse | 5′-ACGACGGCCGCGATCTTATTTC-3′ | |
MT2 | ||
RT primer | Reverse | 5′-pATTAAGGCAGCTGTTGAA-3′ |
1st PCR | Forward | 5′-CCGAAGGAAGGCAAAGGCTGAGAG-3′ |
Reverse | 5′-GGTGCTGGCTGTCTGGATGAAGGT-3′ | |
2nd PCR | Forward | 5′-CCAGAAGGGCTTTTTGTCACCAG-3′ |
Reverse | 5′-TGACTGCAGGCTCGGTGGTAGGT-3′ | |
ORF cloning | ||
MT1 | Forward | 5′-GCGCGGGGCTACAGGATGATG-3′ |
Reverse | 5′-CAGGAGGTCCGCTCCAACACTATG-3′ | |
MT2 | Forward | 5′-GGAGCGCCCCCAAGCAGT-3′ |
Reverse | 5′-AGGTCAAGGTGGCAGGGAAAATG-3′ | |
Plasmid construction | ||
MT1 | Forward | 5′-CACCATGAAGGGCAATGTCAGC-3′ |
Reverse | 5′-TTAAACAGAGTCCACCTTTATTAAATT-3′ | |
MT2 | Forward | 5′-CACCATGCCTGACAACAGCTCCAT-3′ |
Reverse | 5′-CTAGAGAGCACCTTCCTGGACAG-3′ | |