Nucleotide sequence of degenerate and non-degenerate primers used for conventional and quantitative PCR amplification of arginine kinase, V-type H+-ATPase B-subunit, Na+/K+/2Cl-cotransporter, and Na+/K+-ATPase α-subunit cDNAs from gills of Chasmagnathus granulatus
| Name . | Sequence (5′→3′) . | Target cDNA . | ||
|---|---|---|---|---|
| Primers used in initial amplification by conventional PCR | ||||
| AKF51 | CGC TGA GTC TAA GAA GGG ATT | Arginine kinase | ||
| AKR31 | GAT ACC GTC CTG CAT CTC CTT | |||
| HATF2 | GCN ATG GGN GTN AAY ATG GA | V-type H+-ATPase B-subunit | ||
| HATR4 | TGN GTD ATR TCR TCG TTN GG | |||
| COTFX | TNA AYA THT GGG GNG TNA TG | Na+/K+/2Cl- cotransporter | ||
| COTRX | CCR TCR TCR TAN ARC CAC CA | |||
| NAK10F | ATG ACI GTI GCI CAY ATG TGG | Na+/K+-ATPase α-subunit | ||
| NAK16R | GGR TGR TCI CCI GTI ACC AT | |||
| Species-specific primers used in quantitative PCR | ||||
| CgAKF1 | GTT TCA AGC AGA CCG ACA AG | Arginine kinase | ||
| CgAKR2 | CTT CGT TGC ACC ATA CCA G | |||
| CgHATF1 | CCG ATT CTT CAA GCA GGA C | V-type H+-ATPase B-subunit | ||
| CgHATR2 | AAC CAG GGA AAC CAC GAC | |||
| CgCOTF1 | TGG CTC GCA GAT TGA CTT | Na+/K+/2Cl- cotransporter | ||
| CgCOTR2 | TGA AGC ATC CCT CAG TGT AA | |||
| CgNAKF1 | TCC CTT CAA CTC CAC CAA | Na+/K+-ATPase α-subunit | ||
| CgNAKR2 | ATA CCA GCA GAA CGG CAC | |||
| Name . | Sequence (5′→3′) . | Target cDNA . | ||
|---|---|---|---|---|
| Primers used in initial amplification by conventional PCR | ||||
| AKF51 | CGC TGA GTC TAA GAA GGG ATT | Arginine kinase | ||
| AKR31 | GAT ACC GTC CTG CAT CTC CTT | |||
| HATF2 | GCN ATG GGN GTN AAY ATG GA | V-type H+-ATPase B-subunit | ||
| HATR4 | TGN GTD ATR TCR TCG TTN GG | |||
| COTFX | TNA AYA THT GGG GNG TNA TG | Na+/K+/2Cl- cotransporter | ||
| COTRX | CCR TCR TCR TAN ARC CAC CA | |||
| NAK10F | ATG ACI GTI GCI CAY ATG TGG | Na+/K+-ATPase α-subunit | ||
| NAK16R | GGR TGR TCI CCI GTI ACC AT | |||
| Species-specific primers used in quantitative PCR | ||||
| CgAKF1 | GTT TCA AGC AGA CCG ACA AG | Arginine kinase | ||
| CgAKR2 | CTT CGT TGC ACC ATA CCA G | |||
| CgHATF1 | CCG ATT CTT CAA GCA GGA C | V-type H+-ATPase B-subunit | ||
| CgHATR2 | AAC CAG GGA AAC CAC GAC | |||
| CgCOTF1 | TGG CTC GCA GAT TGA CTT | Na+/K+/2Cl- cotransporter | ||
| CgCOTR2 | TGA AGC ATC CCT CAG TGT AA | |||
| CgNAKF1 | TCC CTT CAA CTC CAC CAA | Na+/K+-ATPase α-subunit | ||
| CgNAKR2 | ATA CCA GCA GAA CGG CAC | |||
D=A/G/T; H=A/C/T; I=A/C/G/T; N=A/C/G/T; R=A/G; Y=C/T.