Table 2.
Primers used for RT-PCR and multi-cell RT-PCR
| Gene | Direction | Primer sequences | Product size (bp) |
|---|---|---|---|
| RT-PCR | |||
| MT1 | Forward | 5′-ATGGCCCTGGCTGTGCTGCGGTAAG-3′ | 316 |
| Reverse | 5′-TAAGTATAGACGTCAGCGCCAAGGGAAATG-3′ | ||
| MT2 | Forward | 5′-GGAGCGCCCCCAAGCAGTG-3′ | 390 |
| Reverse | 5′-GGATCTCCCCAAGTACCCAACCGTCAT-3′ | ||
| β-Actin | Forward | 5′-GTCCACACCCGCCACCAGT-3′ | 496 |
| Reverse | 5′-CGTCTCCGGAGTCCATCACAAT-3′ | ||
| mGAPDH | Forward | 5′-TGAAGGTCGGTGTGAACGGATTTG-3′ | 359 |
| Reverse | 5′-GGCGGAGATGATGACCCTTTTG-3′ | ||
| Multi-cell RT-PCR | |||
| MT1 | |||
| 1st PCR | Forward | 5′-ATGGCCCTGGCTGTGCTGCGGTAAG-3′ | 442 |
| Reverse | 5′-TGTGGCAAATGTAGCAGTAGCGGTTCA-3′ | ||
| 2nd PCR | Forward | 5′-CAGGCGGCGGGGAGGAAATAAG-3′ | 231 |
| Reverse | 5′-TAAGTATAGACGTCAGCGCCAAGGGAAATG-3′ | ||
| MT2 | |||
| 1st PCR | Forward | 5′-GGAGCGCCCCCAAGCAGTG-3′ | 390 |
| Reverse | 5′-GGATCTCCCCAAGTACCCAACCGTCAT-3′ | ||
| 2nd PCR | Forward | 5′-CGGGCTGCAGCGTCACCAT-3′ | 275 |
| Reverse | 5′-GTCAGCCAAGGCCAGATTCACC-3′ | ||
| GnRH | Forward | 5′-ACTGATGGCCGCTGTTGTTCT-3′ | 256 |
| Reverse | 5′-CTTCTTCTGCCCAGCTTCCTCTTCA-3′ | ||
Forward and reverse primers were designed on different exons to distinguish the amplification of cDNA templates from that of contaminating genomic DNA. To design the PCR primers, we referred to the following nucleotide sequences: AB377274 for MT1, AB377275 for MT2, NM_031144 for β-actin, NM_008084 for mGAPDH, and M12579 for GnRH