Table 2.
Primers | |||
Assay | Primer | Position | Sequence |
RACE | 5'RACE nested | 48852556 | CCTCTCACAGCTCATGTCTGAT |
RACE | 5'RACE | 48852500 | ACCACATCCAGGCACTGTTCTTC |
RACE | 3'RACE | 48852519 | GAACAGTGCCTGGATGTGGTGAC |
PCR/Real-time | uPAR Fwd E1 | 48866085 | CTGCTGCTGCTCCACACCT |
PCR/Real-time | uPAR Rev E7 | 48845002 | TGGGTGGTTACAGCCACTTT |
PCR/Real-time | uPAR Rev E7b | 48842344 | ATCACATCTGCTGGGGCTAC |
PCR/Real-time | uPAR Rev E2 | 48863624 | ACTCTTCCACACGGCAATCCC |
PCR/Real-time | uPAR Rev E7b (long UTR) |
48842223 | GCCTGTCCTTCCATGATGTT |
Probes | |||
Assay | TaqMan Probe | Location | Sequence ('denotes exon boundary) |
Real-time PCR | Exon 1–2 | Exon1–2 | CCAGCCTCTTGGGGCCTGCGGT |
Real-time PCR | Exon 5 del | Exon4–6 | AGCTCCAGGA'CTTCTTCACCTTCC |
Real-time PCR | Exon 3 del | Exon 2–4 | CAGCCCGGC'CTTCCCACAAGCG |
Real-time PCR | Exon 6 del +E7 | Exon 5–7 | TTCGGTT'TTGGGCCCTCGTTGCAT |
Real-time PCR | Exon 6 del +E7b | Exon 5–7b | TTCCCCAGAGTGAGCGTT'TTGGGC |
Real-time PCR | Exon 5+6 del +E7 | Exon 4–7 | TCCAGGAAGGTGAAGAAG'AACCGAAA |
Real-time PCR | Exon 5+6 del; E7b | Exon 4–7b | CCCAGAGTGAGCGTT'CTTCTTCACC |
Real-time PCR | Exon 4+5 del | Exon 3–6 | TGCAACCAGGGCAACTCTG'TCCT |
Primers listed for RACE were used with RACE-specific primers from GeneRacer kit (Invitrogen). Other primers were used to amplify full-length uPAR for both standard PCR and real-time assays. All TaqMan probes were labelled with Fam and TAMRA and were used with appropriate primers. Primer location is listed relative to chromosome 19 reference sequence for the start of the primer.