TABLE 1.
Primers used for Cloning and RT-PCR
| Oligo ID | 5′-3′ sequence | Annealing (°C) | Comments |
|---|---|---|---|
| mERαUS | AAGGGCAGTCACAATGAACC | 59 | Will amplify mouse ERα (NM_007956) cDNA sequence of 155 bp |
| mERαDS | GCCAGGTCATTCTCCACATT | ||
| mERβUS | GGGTGAAGGAGCTACTGCTG | 59 | Will amplify mouse ERβ transcript variants 1 and 2 (NM_207707 and NM_010157) cDNA sequence of 576 and 522 bp, respectively |
| mERβDS | GTGTCAGCTTCCGGCTACTC | ||
| mGAPDHUS | TGACATCAAGAAGGTGGTGAAG | 55 | Will amplify mouse (NM_008084) glyceraldehyde 3 phosphate dehydrogenase cDNA sequence of 243 bp |
| mGAPDHDS | TCTTACTCCTTGGAGGCCATGT | ||
| mCK19US | GAGATCATGGCCGAGAAGAA | 55 | Will amplify mouse cytokeratin 19 (NM_008471.2) cDNA sequence of 72 bp |
| mCK19DS | GGTGTTCAGCTCCTCAATCC | ||
| mVimentinUS | GTGGCTCCGGCACATCGAGC | 56 | Will amplify mouse vimentin (NM_011701.4) cDNA sequence of 226 bp |
| mVimentinDS | GCGTCGGCCAGCGAGAAGTC | ||
| mCYP2E1US | GTGTTCCGAGGATATGTCATC | 56 | Will amplify mouse CYP2E1 (NM_021282.2) cDNA sequence of 223 bp |
| mCYP2E1DS | AAAGCAGAAACAGTTCCATGC | ||
| mERaCloningDSHindIII | CCTGGAAGCTTTCAGATCGTGTTGGGG | 72 | Will amplify mouse ERα transcript variants 1, 2 and 3 (NM_007956.5, NM_001302531.1, NM_001302532.1) cDNA sequence of 1829 bp (2-step PCR) |
| mERaCloningUSEcoRI | CGCCGAATTCCACTTACCATGACCATG | ||
| mERβUScloning | TCCGCCTTAAGCCTGGCCGTCACTG | 65 | Will amplify mouse ERβ transcript variants 1 and 2 (NM_207707 and NM_010157) cDNA sequence of 1744 and 1690 bp, respectively |
| mERβDScloning | TCCGCCTTAAGCCTGGCCGTCACTG |