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Table 1.

Oligonucleotide primers used for cDNA cloning, RACE, real-time PCR and probe construction for in situ hybridization and Northern blotting

Primer namePrimer sequenceUsesProduct sizePrimer 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 namePrimer sequenceUsesProduct sizePrimer 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)

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