Table 1.
Primer and probe sequences and concenrations used in real time PCR
| Genes | Sequence(5′–3′) | Nucleotide position | Final concentraion (nm) |
|---|---|---|---|
| 18S | |||
| Forward primer | 5′-CGGCTACCACATCCAAGGAA-3′ | N/A | 40 |
| Reverse primer | 5′-GCTGGAATTACGCGGCT-3′ | N/A | 20 |
| Taqman probe | 5′TGCTGGCACCAGACTTGCCCTC-3′ | N/A | 50 |
| AQP 1 | |||
| Forward primer | 5′CCATCGTCGCCACTGTCAT-3′ | 234–342 | 300 |
| Reverse primer | 5′-GAGGCCAAGCGAGTTGTCA-3′ | 388-370 | 300 |
| Taqman probe | 5′-TCTCGGGCATCACTCCTCTCTGC-3′ | 345–368 | 75 |
| AQP 3 | |||
| Forward primer | 5′-GGGTGCCCATTGTCTCTCC-3′ | 500–518 | 900 |
| Reverse primer | 5′-AGCCCGATCATGAGCTGGTACA-3′ | 568-547 | 900 |
| Taqman probe | 5′-TCCTGGTTCCATCGCGGGTG-3′ | 521–540 | 75 |
| AQP 4 | |||
| Forward primer | 5′-AGAATTTCTGGCCATGCTTATTTT-3′ | 135–158 | 300 |
| Reverse primer | 5′-TCTGCTCCACCCCAGTTGA-3′ | 203–185 | 900 |
| Taqman probe | 5′-TCCTGCTCAGCCTGGATCCACC-3′ | 161–183 | 75 |
| AQP 5 | |||
| Forward primer | 5′-CGCCGCAATCCTCTATGG-3′ | 315–332 | 300 |
| Reverse primer | 5′-GCTGGAATTACCGCGGCT-3′ | 396–376 | 900 |
| Taqman probe | 5′-CAATGCCCGAAGCAATCTGGCTG-3′ | 345–367 | 100 |
For each AQP TaqMan probe, FAM (6-carboxy fluorescein) was attached at the 5′ end and TAMRA (6-carboxy-tetramethyIrhodamine) was attached at the 3′ end. Accession numbers (GenBank/EMBL) are as follows: AQP1-AF00937, AQP3-AF123316. VIC was used for 18S.