Table 1.
Oligonucleotide primer sequences. Numbering of nucleotide position corresponds to ERαL (GenBank: HM641843) except for the ERαS-specific primers Q1 and Q2, indicated by asterisk (*), where numbering corresponds to position in ERαS (GenBank: AY571785). With the exception of primer set Q5/Q6 (Greytak and Callard, 2007; Greytak et al., 2010), all other primers were designed in this study. See Supplementary Table 2 for additional primers.
| Primer | Exon Position | Nucleotide Position | Sequence 5′-3′ | |
|---|---|---|---|---|
| PCR cloning | 1 (F) | Exon 2a | 162–186 | GCAAAATKGGAKARRSRAGTCTAAATC |
| 2 (R) | Exon 2b | 294–314 | GGAGAGGGTCTCCAGCTCTGA | |
| 3 (F) | Exon 1 | 1–23 | GCCCCGAGGATGATTCATGTATAA | |
| 4 (F) | Exon 5a | 1138–1157 | GGGACAGATCTTCAGTGGCC | |
| 5 (F) | Exon 5b | 1286–1305 | CAAGGAGCTGGTCCACATGA | |
| 6 (R) | Exon 5b | 1219–1238 | TCGGCTAAGTTTTTGACGGG | |
| 7 (R) | Exon 5b | 1230–1255 | GTGACCTCGGTGTAGGGTCGGCTAAG | |
| 8 (R) | Exon 8 | 1674–1692 | TGAGAGCGTCGGTGATGGT | |
| 9 (R) | Exon 9 | 2074–2092 | TTCAGGCCTCCGACTCACA | |
| qPCR | Q1 (F) | Exons 1/2b | 8–33* | GATGATTCATGTATAAAGGGCAGAAC |
| Q2 (R) | Exon 2b | 94–114* | GGAGAGGGTCTCCAGCTCTGA | |
| Q3 (F) | Exon 2a | 110–132 | GAAAAGGGTGAGACCCAGAGAGA | |
| Q4 (R) | Exon 2a | 191–210 | CACGGCAGCACTGATAAGCA | |
| Q5 (F) | Exon 7 | 1540–1558 | GCATGCTCAAGCTCAAACC | |
| Q6 (R) | Exon 8 | 1611–1627 | GTGCCGGTGCAGAAAGA | |
| Expression vector cloning | SF | Exon 2a | 357–374 | CACCATGTACCCTGAAGAGAGC |
| LF | Exon 2b | 216–233 | CACCATGTTGCTCAGGCAGAAC | |
| R | Exon 9 | 2060–2078 | TCACAGGACCTGGGCACAG |