Table 1 Primers used for cloning, sequencing, transcriptional analysis and probe synthesis
From: Morphological and molecular characterization of developing vertebral fusions using a teleost model
Gene | Orientation | Genbank | Use | Sequence (5'-3') |
|---|---|---|---|---|
Extracellular Matrix constituents | ||||
Col1a1 | Forward | FJ195608 | RT | AGAGAGGAGTCATGGGACCCGT |
| Â | Reverse | Â | RT | GGGTCCTGGAAGTCCCTGGAAT |
| Â | Forward | Â | ISH | TAGCCGTGGTTTCCCTGGTT |
| Â | Reverse | Â | ISH | CCGGGAGGTCCAAATCTACC |
Col2a1 | Forward | FJ195613 | RT | TGGTCGTTCTGGAGAGACT |
| Â | Reverse | Â | RT | CCTCATGTACCTCAAGGGAT |
| Â | Forward | Â | ISH | GCTGGCGAGACAGGAGAGA |
| Â | Reverse | Â | ISH | GCCTCATCAGCCCTCATGTA |
Col10a1 | Forward | EG837148 | RT | TGGTGCTCTTTGACTGCCTGTAA |
| Â | Reverse | Â | RT | CATCCTGTGTGTTGCAATATCACA |
| Â | Forward | Â | ISH | AACAAGGGCTTCTTGGATCA |
| Â | Reverse | Â | ISH | CATAATGCATCCTCAGGCAT |
Alp | Forward | FJ195609 | RT | CTAGTTTGGGTCGTGGTATGT |
| Â | Reverse | Â | RT | TGAGGGCATTCTTCAAAGTA |
Osteocalcin | Forward | FJ195616 | RT | GTGAACCAACAGCAAAGAGA |
| Â | Reverse | Â | RT | CCAGGTCCTTCTTAACAAACA |
| Â | Forward | Â | ISH | CTCATACTTGTTGATCGTCCAG |
| Â | Reverse | Â | ISH | TCTTTCTCTCTCGCTCTCCC |
Osteonectin | Forward | FJ195614 | RT | ATTACTGAGGAGGAGCCCATCATT |
| Â | Reverse | Â | RT | CCTCATCCACCTCACACACCTT |
| Â | Forward | Â | ISH | CTGAACGATGAGGGTGTGGA |
| Â | Reverse | Â | ISH | CGAGTGGTGCAGTGCTCCAT |
Mmp9 | Forward | CA342769 | RT | AGTCTACGGTAGCAGCAATGAAGGC |
| Â | Reverse | Â | RT | CGTCAAAGGTCTGGTAGGAGCGTAT |
Mmp13 | Forward | DW539943 | RT | TGATGTCCAAGTCAGCCGCTTC |
| Â | Reverse | Â | RT | TGGTCTGCCACTTGCGATTGTC |
Cathepsin K | Forward | NM_001140399 | RT | ATGACCAACGCCTTCGAGTAC |
| Â | Reverse | Â | RT | AAGGTGGAGAGGGTGGCATC |
Transcription factors | Â | Â | Â | Â |
Sox9 | Forward | EU344852 | RT | CCTGCAAACAAGACAAGGT |
| Â | Reverse | Â | RT | GGGTCGAGTAGATTCATACGA |
| Â | Forward | Â | ISH | GGGGATACTATTTGACTGGATC |
| Â | Reverse | Â | ISH | TCTGTCTTGATGTGTGTGGG |
Runx2 | Forward | FJ195615 | RT/ISH | CCACCAGGGACAGACACAGAT |
| Â | Reverse | Â | RT/ISH | GAACGGACTGAGATCTGACGAA |
Osterix | Forward | FJ195612 | RT | TCCCATAGACTTTCCCACA |
| Â | Reverse | Â | RT | TGCCTCAGGACATGTACAA |
Mef2c | Forward | GU252207 | RT | CACCGTAACTCGCCTGGTCT |
| Â | Reverse | Â | RT | GCTTGCGGTTGCTGTTCATA |
| Â | Forward | Â | ISH | GACAGAGACTGTGTGGTGTTCCCT |
| Â | Reverse | Â | ISH | AGGTGGAGGGAGCTACCACTGTTA |
Signalling molecules | Â | Â | Â | Â |
Bmp4 | Forward | FJ195610 | RT | TCAAGTTGCCCATAGTCAGT |
| Â | Reverse | Â | RT | CACCTGAACTCTACCAACCA |
Bmp2 | Forward | BT059611 | RT | ATGTGGTATTGCACCCATT |
| Â | Reverse | Â | RT | ATGGACAGTTTCCCAATGA |
Shh* | Forward | AY370830 | RT | CCGGCTCATGACTCAGAGATG |
| Â | Reverse | Â | RT | TATCCCTGGCCACTGGTTCA |
Ihh | Forward | FJ195617 | RT | CAGATGACCCACTGGACTGAT |
| Â | Reverse | Â | RT | GCTTGGTTGGGAGATATGCA |
Housekeeping gene | Â | Â | Â | Â |
Ef1a ** | Forward | DQ834870 | RT | CACCACCGGCCATCTGATCTACAA |
| Â | Reverse | Â | RT | TCAGCAGCCTCCTTCTCGAACTTC |