Primers used for PCR and sequencing of 5′ flanking region of the FOXC2 gene promoter
| Primers (5′–3′) | 5′ position of each primer | Buffer and DNA polymerase for PCR |
|---|---|---|
| PCR | ||
| pr1F CCAAACCCCACAAAACTGCTCGCAGCGACG | −1248 | Advantage GC genomic polymerase mix |
| pr1R GTAATTCTGCTCGCTCAGGTAGGGCACCAC | +72 | |
| Sequencing | ||
| pr1F CCAAACCCCACAAAACTGCTCGCAGCGACG | −1248 | |
| 1RN1 TCAGGTAGGGCACCACTCC | +58 | |
| pr2F GGCCCCCATAATTAGGAAA | −938 | |
| pr2R ATCTCTCCCAAAGACCTTG | −841 | |
| pr3F TCTTAGAGCCGACGGATTC | −645 | |
| pr3R CCCGGAACTTTGAGCCAAT | −560 | |
| pr4F GTCCTGGAGCCAGCGAGGA | −283 | |
| pr4R TTTCAGCGGACCGGGCGGA | −198 |
The 5′ positions are shown by defining the translation start site as +1. The 5′ sequences were determined based on the comparison between the human draft sequence for the FOXC2 gene on chromosome 16 (GenBank accession no. NT_024788) and the human FOXC2 cDNA sequence (no. NM_005251). One large fragment was amplified by PCR with primers indicated as PCR and each region was sequenced with ones indicated as sequencing as described in research design and methods. F, forward; R, reverse; N, nested.