Sequences of oligonucleotides used for EMSAs, site-directed mutagenesis, and PCR cloning For oligonucleotides used as EMSA probes or competitors, or in site-directed mutagenesis, only the top strands are shown. Where applicable, the engineered restriction enzyme sites are underlined, and the corresponding restriction enzymes are given in parentheses after the sequences.
Oligonucleotide | Sequence (5′-3′) | Purpose |
|---|---|---|
| HNF-4α consensus | GATCAGGTCTCACAGGTCAAAGGTCACCCTGGGA | EMSA probe and competitor |
| hOCT1 WT A | GATCCTATGGAGCCCTATTGACCCTGGAGT | EMSA competitor |
| hOCT1 WT B | GATCTCCTGTTGATCTCTTGTCCTTCCCTT | EMSA competitor |
| hOCT1 mut A | GATCCTATGGACCCCTATTCACCCTGGAGT | EMSA competitor |
| hOCT1 mut B | GATCTCCTGTTCATCTCTTCTCCTTCCCTT | EMSA competitor |
| hOCT1(–1479/–1441) WT | GATCCTATGGAGCCCTATTGACCCTGGAGTCCTGTTGATCT CTTGTCCTTCCCTT | EMSA probe |
| hOCT1(–1479/–1441) mut A+B | GATCCTATGGACCCCTATTCACCCTGGAGTCCTGTTCATCT CTTCTCCTTCCCTT | EMSA probe |
| hOCT1(–1479/–1441) mut A | GATCCTATGGACCCCTATTCACCCTGGAGTCCTGTTGATCT CTTGTCCTTCCCTT | EMSA probe |
| hOCT1(–1479/–1441) mut B | GATCCTATGGAGCCCTATTGACCCTGGAGTCCTGTTCATCT CTTCTCCTTCCCTT | EMSA probe |
| hOCT1(–2620/+116) mut A+B | CAGCTATGGACCCCTATTCACCCTGGAGTCCTGTTCATCTC TTCTCCTTCCCTTC | Site-directed mutagenesis |
| hOCT1(–2620/+116) mut A | CAGCTATGGACCCCTATTCACCCTGGAGTCCTGTTGATCTC TTGTCCTTCCCTTC | Site-directed mutagenesis |
| hOCT1(–2620/+116) mut B | CAGCTATGGAGCCCTATTGACCCTGGAGTCCTGTTCATCTC TTCTCCTTCCCTTC | Site-directed mutagenesis |
| hOCT1(+116)rev | CATTGCACCTGGCCACTCGAGCCCAGAGCAGGTCTGGCCA (Xho I) | hOCT1(–3852/+116), (–2620/+116), (–1356/+116), (–880/+116) cloning |
| hOCT1(–3852)for | CTGGTTTTGGGGTCCTGTGGTACCCCATGCCTGCTGCTGT (Kpn I) | hOCT1(–3852/+116) cloning |
| hOCT1(–2620)for | TTCCCCCTCTGAAGAGGTACCCCTAACTGCTGTTAGGGTA (Kpn I) | hOCT1(–2620/+116) cloning |
| hOCT1(–1356)for | TCTTGTTCAAGTGAAAGCCAAATGGTACCCTTACATTAGTC TATTAATGTCAACCCCA (Kpn I) | hOCT1(–1356/+116) cloning |
| hOCT1(–880)for | CAGTAGTCATCACTACATGTTATAATGGTACCTCTTAGTAAT (Kpn I) | hOCT1(–880/+116) cloning |