Oligonucleotide primers used for cDNA cloning, RACE, real-time PCR and probe construction for in situ hybridization and Northern blotting
| Primer name . | Primer sequence . | Uses . | Product size . | Primer position . |
|---|---|---|---|---|
| DNHE2-F | 5′-AAYGAYGSIGTIACIGTIGT-3′ | Cloning of partial NHE2 cDNA | – | 730–749 |
| DNHE2-R | 5′-GGICKIATIGTIATICCYTG-3′ | Cloning of partial NHE2 cDNA | 704 | 1414–1433 |
| 3NHE2F1 | 5′-TCAGGGAGATAGAACCCCTCT-3′ | Outer 3′ RACE | 911–931 | |
| 3NHE2F2 | 5′-TATGGCCATTGTGACCTGTG–3′ | Inner 3′ RACE | 993–1012 | |
| 5NHE2R1 | 5′-GACCACACCCAGGAAGAAGA-3′ | Outer 5′ RACE | 1118–1137 | |
| 5NHE2R2 | 5′-AGACCAGGGCAAAGGAGATT-3′ | Inner 5′ RACE | 1314–1333 | |
| QNHE2-F | 5′-TGTGCCCTGACCATGAAGTA-3′ | NHE2 RT-PCR | – | 1009–1028 |
| QNHE2-R | 5′-CCCAGTTCCACTCGTGTTCT-3′ | NHE2 RT-PCR | 160 | 1149–1168 |
| NHE2-F | 5′-CGGGCAGAGTCTAGCATTGT-3′ | In situ hybridization | – | 1630–1649 |
| NHE2-R | 5′-TGTCGTCACGGTCACCAG-3′ | In situ hybridization | 811 | 2423–2440 |
| DNHE3-F | 5′-CTTCATGTTYCTKGGHATCTCKGC-3′ | Cloning of partial NHE3 cDNA | – | 511–534 |
| DNHE3-R | 5′-CATGGTCCKSTGGAARATYTC-3′ | Cloning of partial NHE3 cDNA | 1167 | 1657–1677 |
| 3NHE3F1 | 5′-ATCCTCCTCACACTCCTCTTCATCTT-3′ | Outer 3′ RACE | 1162–1187 | |
| 3NHE3F2 | 5′-CAGGTGGTGATGAGCTACGGT-3′ | Inner 3′ RACE | 1267–1287 | |
| 5NHE3R1 | 5′-TTATTCCCTGCAGCATGACA-3′ | Outer 5′ RACE | 1395–1414 | |
| 5NHE3R2 | 5′-GCCGAGGAAGACAAAGATGA-3′ | Inner 5′ RACE | 1097–1116 | |
| NHE3-F | 5′-GCTCCCTGGTTGGTATTATC-3′ | Northern blot probe | – | 833–852 |
| NHE3-R | 5′-AACCAGCACAACCACCTCTC-3′ | Northern blot probe | 772 | 1585–1604 |
| QNHE3-F | 5′-AGAGCAGCCGTGACAGAACT-3′ | NHE3 RT-PCR | – | 1450–1469 |
| QNHE3-R | 5′-AACCAGCACAACCACCTCTC-3′ | NHE3 RT-PCR | 155 | 1585–1604 |
| Actin-F | 5′-CCAACAGATGTGGATCAGCAA-3′ | RT-PCR control | – | 1056–1076 |
| Actin-R | 5′-GGTGGCAGAGCTGAAGTGGTA-3′ | RT-PCR control | 138 | 1173–1193 |
| Primer name . | Primer sequence . | Uses . | Product size . | Primer position . |
|---|---|---|---|---|
| DNHE2-F | 5′-AAYGAYGSIGTIACIGTIGT-3′ | Cloning of partial NHE2 cDNA | – | 730–749 |
| DNHE2-R | 5′-GGICKIATIGTIATICCYTG-3′ | Cloning of partial NHE2 cDNA | 704 | 1414–1433 |
| 3NHE2F1 | 5′-TCAGGGAGATAGAACCCCTCT-3′ | Outer 3′ RACE | 911–931 | |
| 3NHE2F2 | 5′-TATGGCCATTGTGACCTGTG–3′ | Inner 3′ RACE | 993–1012 | |
| 5NHE2R1 | 5′-GACCACACCCAGGAAGAAGA-3′ | Outer 5′ RACE | 1118–1137 | |
| 5NHE2R2 | 5′-AGACCAGGGCAAAGGAGATT-3′ | Inner 5′ RACE | 1314–1333 | |
| QNHE2-F | 5′-TGTGCCCTGACCATGAAGTA-3′ | NHE2 RT-PCR | – | 1009–1028 |
| QNHE2-R | 5′-CCCAGTTCCACTCGTGTTCT-3′ | NHE2 RT-PCR | 160 | 1149–1168 |
| NHE2-F | 5′-CGGGCAGAGTCTAGCATTGT-3′ | In situ hybridization | – | 1630–1649 |
| NHE2-R | 5′-TGTCGTCACGGTCACCAG-3′ | In situ hybridization | 811 | 2423–2440 |
| DNHE3-F | 5′-CTTCATGTTYCTKGGHATCTCKGC-3′ | Cloning of partial NHE3 cDNA | – | 511–534 |
| DNHE3-R | 5′-CATGGTCCKSTGGAARATYTC-3′ | Cloning of partial NHE3 cDNA | 1167 | 1657–1677 |
| 3NHE3F1 | 5′-ATCCTCCTCACACTCCTCTTCATCTT-3′ | Outer 3′ RACE | 1162–1187 | |
| 3NHE3F2 | 5′-CAGGTGGTGATGAGCTACGGT-3′ | Inner 3′ RACE | 1267–1287 | |
| 5NHE3R1 | 5′-TTATTCCCTGCAGCATGACA-3′ | Outer 5′ RACE | 1395–1414 | |
| 5NHE3R2 | 5′-GCCGAGGAAGACAAAGATGA-3′ | Inner 5′ RACE | 1097–1116 | |
| NHE3-F | 5′-GCTCCCTGGTTGGTATTATC-3′ | Northern blot probe | – | 833–852 |
| NHE3-R | 5′-AACCAGCACAACCACCTCTC-3′ | Northern blot probe | 772 | 1585–1604 |
| QNHE3-F | 5′-AGAGCAGCCGTGACAGAACT-3′ | NHE3 RT-PCR | – | 1450–1469 |
| QNHE3-R | 5′-AACCAGCACAACCACCTCTC-3′ | NHE3 RT-PCR | 155 | 1585–1604 |
| Actin-F | 5′-CCAACAGATGTGGATCAGCAA-3′ | RT-PCR control | – | 1056–1076 |
| Actin-R | 5′-GGTGGCAGAGCTGAAGTGGTA-3′ | RT-PCR control | 138 | 1173–1193 |
The expected PCR product sizes and the positions of the PCR primers were deduced on the basis of the GenBank accession numbers EF446605 (NHE2)and EF446606 (NHE3)