Table 2.
Primers Used to Sequence 24 Exons of TRIOBP and for Cloning Full-Length Isoforms and RT-PCR[Note]
|
Primer Sequence(5′→3′) |
|||
| Use for Primerand Exon(s) or Isoform | Forward | Reverse | Product(bp) |
| Sequencing of exons: | |||
| 1 | GAGGGATCCGGGGAGGAA | GGAGTGGAGCTGGGTCAGGA | 695 |
| 2 | ATTTCCTTGGTCCTTGTCTGGAAAC | GAAGTCCAGCATCTGGGTTCAACT | 382 |
| 3 | CCGAGGAGATGCCTGGAGACT | ATGCCAGCAGCCATTAGAGACAG | 356 |
| 4 | CTCTCACCATTGGCTCCCTTT | CTGTGCAGAAATCCCCACTCTC | 523 |
| 5 | TCTGCCCCTCTCATTTGGAGAGTAT | TAACACTCAGGCCACCAAAGTAAGC | 368 |
| 6 | AGTGAGACCTGTAGGAGGAGGGTGT | CCACACACACACACAAAACAGAAAA | 3,641 |
| 7 | CTGTTGGAGGTGGGAGCAGAG | CATAGAGGACCTTCCTGTCCAAGC | 300 |
| 8a | CAGCCAGGTCCCCAGAGAGAG | GCTCTCCACATCCCCACCAG | 383 |
| 8b | CCAGGGCCCTCATAGACACCTAGAA | CCCAGACCCCCTCTCCTGGT | 416 |
| 8c | AGCTTGACTGGAGGGATCTGCTT | AGTGGGGCTGACTTTCTGGCTAC | 489 |
| 9 | ATAGAATGGATGGATGAGGTGGACA | CTAGGGCTCTTCTGAGGAATGAAGC | 250 |
| 10 | AATGGGCAGGGGAGGCTGTAG | CTCCCTCCCATCCCCGTCTCTAT | 281 |
| 10A | CCGCTGTTTGTCTCGGCTTC | CTCTCGCCTTCGGGAAATGAG | 382 |
| 11 | CCACAGGACTTGGGGACAGG | AGCAGCACAAGCCAGCTTCC | 203 |
| 12 and 13 | AGGCTCAGAGAGGAAATGGAG | GTGGATCTGGAAGCCATAGTTG | 463 |
| 14 | CACTTAGCATATCTGGGGGAGA | CTCTTCCCATGTTGTTGACAGA | 444 |
| 15a | AGGGGTGAGGACCCACCTGA | CTGTCTGTGGCCTCAAACCACTTG | 339 |
| 15b | ACCCCAGCCCGCACTCCTGAC | CAGGGGTGAGCTCCCCATCACTGTA | 448 |
| 16 | TCTCAAACCCTGAGGCATTC | GGGGAGAGTAAATGAGATGACG | 402 |
| 17 | CTCCTCTGGAGGCTGGGGAAG | CCAGTTTGGCTAGGAGGAACCTCAG | 297 |
| 18 | GTGCTCTTTGCATAGCACTCCT | ACCTTCTGCTTCTGTTTCTTGC | 416 |
| 19 and 20 | GAGTCCTGGGATGCTTTAAGTC | CTTTCCTGCCACAGGTGATAC | 540 |
| 21 | AGATAGGGGCTGCATTCTAGG | AGAGCCTGAGGAAGGATGAAG | 318 |
| 22 | AGGGATGAAGCGAGGTATCTG | CTTTTTACAGAACGGGAATTGG | 406 |
| 23 | CTGAGCTGCTTGGGAAGGA | GGCCACCAGCTCAGAAGC | 598 |
| Cloning of full-length isoforms: | |||
| TRIOBP-6/3 | TCATAGGAACTGCCCTGGCCTGACT | GCACGTGTGTGCGTATGTGTCTGTG | 7,113 |
| TRIOBP-4 | CCAAGATTGGCCACAAAAGCCTGAT | GTCCTGCTGCCGAGCCTCCTC | 3,901 |
| Triobp-1 | GGAAGGGAAGGGCTGGAGCTACG | GGGTGACTCACGGAACAGAAGAG AAGTG | 2,492 |
| Triobp-3 | AGTCGCACACTGACCATGCAAATGA | ACGAGCTCAGATCCGCAGTGTCTGT | 6,801 |
| Triobp-4 | AGTCGCACACTGACCATGCAAATGA | GGGAAGTTAAATTGCTTTAATCAGGT ATCACAGGA | 3,740 |
| Triobp-5 | AGTCGCACACTGACCATGCAAATGA | ACGAGCTCAGATCCGCAGTGTCTGT | 6,663 |
| RT-PCR: | |||
| Triobp-3 | ACCTGAGGATGGGGCCTGGAC | CGCTGCAGCTGCTCCATCACC | 1,635 |
| Triobp-4 | CTCAGAGGGTCTGGTCCCCAACAAC | GCTCCTGTGTTTTTGACTACCTGG CTA | 2,870 |
| Triobp-5 | ACCTGAGGATGGGGCCTGGAC | CGCTGCAGCTGCTCCATCACC | 1,496 |
Note.— All the primers were designed using Primer3.