Table A2.
List of primers for replacement constructs and RT-PCR.
| Gene | Primer 1 | Sequence (5'–3') a | Primer 2 | Sequence (5'–3') | Size (kb) | Application |
|---|---|---|---|---|---|---|
| PP104 | paxU1SstIL | CTTGTTGGCgaGCTcCATATGAC | pax66 | CGCGATGGCGTACTGTAGAC | 1.03 | KO construct |
| PP104 | paxRU1HindIIIL | TTTAGTAGAAGCTTGGCC | paxRU1HindIIIR | TCGTTGAAGcTTGCAGTA | 1.16 | KO construct |
| PP105 | paxRBamL | ATTGACGgATCCCGATTATC | paxRBamR | GGATccGAGATGGGTGTATAC | 1.1 | KO construct |
| PP105 | KORH | GGGGTATAaGcTTAACATAGAGCAG | KORHS | GTTACATGCTTCCATTTAAAGTTGGGAGCTGTC | 1.1 | KO construct |
| PP104 & PP105 | paxRU1BamR | CAACGTTGTGGATCCATTCGG | paxRU1BamL | CCCTATCGGGATGCAATTTTCAAAC | 1.1 | KO construct |
| PP107 | pax251Hind | TGCAAGCTTCCGCTATAG | pax186 | AGTCAACACCAAGACAGG | 1.27 | KO construct |
| PP107 | pax175 | TCGACGACTTCGACCAGA | pax183 | ATGTCATCTTCCGCAATC | 1.5 | KO construct |
| PP112 | paxUKOp3 | ACGTTGCTAGTcTAGaTGGAAGC | paxUKOp4 | AGTTCGTaAGCTTGATGTGTTG | 1.5 | KO construct |
| PP112 | paxUKOp1 | GTGATGGATcCCAAAATTCATTGG | paxUKOp2 | GCAAGAaTTCAAATGCCTGGAAG | 1.6 | KO construct |
| paxA | sec23KOp3 | TGGCCGAaTTCCGAGAATAGAGT | sec23KOp4 | AAGAAATACGTGgATCCTGACAG | 1.4 | KO construct |
| paxA | sec23KOp2 | GCTAAAGcTtAACAACTGGACCA | sec23KOp1 | CTCGACAAGcTTAGAAAAGTCAC | 1.5 | KO construct |
| paxB | sec25KOp3/ | CGTTGAATAGCTcTAGATTGAAGG | sec25KOp4 | TGAGCCAAgcTTTGTGTAACTCG | 1.0 | KO construct |
| paxB | sec25KOp1/ | GTGATgGATCCCAAAATTCATTGG | sec25KOp2 | GCAAGaATTCAAATGCCTGGAAG | 1.5 | KO construct |
| paxC | paxCSstR | GTTGAGCTCAATCCACCAACGC | paxCSstL | GAGTGAGCTCTGCTTGGTAGGC | 1.4 | KO construct |
| PP112 | paxU RT-F | TCGTCCTATCTCGCACCTTTC | paxU RT-R | AGAGTCTGTTCGGTTCGATGG | 474 | RT-PCR |
| paxG | paxG RT-F | ACACTGCATCCCTTCTTATCG | paxG RT-R | TATCGAGAAGCTCGGAGCTCT | 528 | RT-PCR |
| paxA | paxA RT-F | CAACCTTTCAGGGTGAGATTC | paxA RT-R | CAGATGAGCAAGCCAAGGCAA | 489 | RT-PCR |
| paxM | paxM RT-F | TCATCGATCAAAGGTTCGGTT | paxM RT-R | AACTCGACCGTAAGCTTGGAA | 301 | RT-PCR |
| paxB | paxB RT-F | GAACTGGTCTACTGTCTGGTC | paxB RT-R | ACGGTCGACGTACCAGAAACA | 504 | RT-PCR |
| paxC | paxC RT-F | ATGATGGTCGACGATATCTCC | paxC RT-R | CAATTGCGAATGCCAGCCAAG | 385 | RT-PCR |
| paxP | paxP RT-F | CCACCTACAAGACCAATGTCA | paxP RT-R | AAGCGAATTGATCATCGCATG | 417 | RT-PCR |
| paxQ | paxQ RT-F | CAGCCTTACAGAGAGATTCGT | paxQ RT-R | GATGTGCGACAACTCTTGCAC | 562 | RT-PCR |
| paxD | paxD RT-F | CAGTCTGGAGCTTATGCCATC | paxD RT-R | CGTCCTTGACGAATGCCTTGA | 456 | RT-PCR |
| PP121 | paxO RT-F | GTGGCTGCTACTAAGCTGGTA | paxO RT-R | CACAGGAAGAAGCGATCTGGT | 456 | RT-PCR |
| PP122 | Pax248 | AGTTCGACAGCGCTTGGGAGA | Pax249 | CAGTGGCTCCTTAACTCTCGT | 592 | RT-PCR |
| tubA | Tub2 RT-F | ACACTCCTGATCTCCAAGATC | Tub2 RT-R | GATGTCGTACAGAGCCTCGTT | 258 | RT-PCR |
a Nucleotides in lower case are mismatches to genomic sequence to introduce sites for restriction enzyme digestion.