Table 1.
Nucleotide base sequences of primers
| Primer sets | Forward | Reverse | Annealing temperature | Elongation time | Lane Number | Expected sizes | |
|---|---|---|---|---|---|---|---|
| For amplification of cDNA | 1 | 5'-GCGGCACCTCGGAAGGC-3' | 5'-CCCCAAACTTTGACTTTTAGC-3' | 60°C | 1 min | Figure 1 A 1 | 834 bp |
| 2 | 5'-GATCGGTTTATCCCTTGTGGTC-3' | 5'-CTTCAGGTTCTTTAAAGTTTGG-3' | 60°C | 1 min | Figure 1 A 2 | 865 bp | |
| 3 | 5'-CTGAAGGGATGTCCAATGC-3' | 5'-ATATTTTATATGATTCTTTTCCTC-3' | 56.1°C | 1 min | Figure 1 A 3 | 850 bp | |
| 4 | 5'-CCAGTCTGTTAACTCCTAGC-3' | 5'-GGATAATGTTCCTCAATATCTTTG-3' | 60°C | 1 min | Figure 1 A 4 | 826 bp | |
| 5 | 5'-ACAGCTTCTAATAAAGAGATAAA AC-3' | 5'-GCCGGCATTTATTATTTTTC-3' | 56.1°C | 1 min | Fig. 1 A 5 | 850 bp | |
| 6 | 5'-GTTTCTCCTCAAGCAAATACAA-3' | 5'-ATTTTTTACTTTGTCCAAAGATTCC-3' | 60°C | 1 min | Figure 1 A 6 | 873 bp | |
| 7 | 5'-CTGATCCTGCAGCAAAGACC-3' | 5'-GAAAAACCAATGTTTTTTCTCTCTC-3' | 59.2°C | 1 min | Figure 1A 7 | 908 bp | |
| 8 | 5'-CATTCTAGTGAAGTGTATAATAA ATCAG-3' | 5'-CTGTCCTAAATCCAGAGAAAGC-3' | 50.8°C | 1 min | Figure 1 A 8 | 919 bp | |
| 9 | 5'-AGTATCACTTAAAGATAATGAAG AAC-3' | 5'-CTTTTAGGATGCCGTCTGG-3' | 50°C | 1 min | Figure 1 A 9 | 887 bp | |
| 10 | 5'-CCCCCAATTAAAAGAAACTTG-3' | 5'-GCCAATTGTATTCCTTCTCC-3' | 53.7°C | 1 min | Figure 1 A 10 | 905 bp | |
| 11 | 5'-CCTCTGCATGTTCTCATAAAC-3' | 5'-GGGTATGCTCTTTGAACAACTAC-3' | 60°C | 1 min | Figure 1 A 11 | 886 bp | |
| 12 | 5'-CATGGAGCAGAACTGGTAGG-3' | 5'-GTGTAAGGTTTAATAATGTCTTCA-3' | 50°C | 1 min | Figure 1 A 12 | 1094 bp | |
| 13 | 5'-CCTATCCCAAGTTTATCAGCC-3' | 5'-CAGACACAAGTTGATGTTCTCC-3' | 60°C | 1 min | Figure 1 A 13 | 959 bp | |
| 14 | 5'-GAAGGCATTTCAGCCACCACG-3' | 5'-CAATCACACTAGAATCATAAAAAGG-3' | 60°C | 1 min | Figure 1 A 14 | 978 bp | |
| For amplification of genomic DNA | exon 1-2 | 5'-GCCCCCTGCCCAGAACCC-3' | 5'-CTTTTCAGCAAGCATGCACAGTTACG-3' | 60°C | 2 min | Figure 1 B 1 | 1193 bp |
| exon 3 | 5'-CTACAGTCAAAATGTCAAGCG-3' | 5'-CACAATTAACAATAGATCTGGGAG-3' | 60°C | 1 min | Figure 1 B 2 | 430 bp | |
| exon 4-7 | 5'-ATAAGAATAAAAACTTCCAGAGAATG-3' | 5'-ATTATCTTTTCATATATTCTTTTTGTC-3' | 60°C | 2 min | Figure 1 B 3 | 1384 bp | |
| exon 8-9 | 5'-GTAGTATATGTGACTTTTGATGTCTG-3' | 5'-GGAAAAGCAATGTATTTTCTCTTT-3' | 60°C | 2 min | Figure 1 B 4 | 615 bp | |
| exon 10 | 5'-CTTTAAATACTGCCTTATGGGCTA-3' | 5'-GTCACCATCCCTAAAACTATATGC-3' | 60°C | 2 min | Figure 1 B 5 | 1311 bp | |
| exon 11-a | 5'-GTCACTTTGTGTCTTCATGC-3' | 5'-GGATAATGTTCCTCAATATCTTTG-3' | 56.4°C | 2 min | Figure 1 B 6 | 1246 bp | |
| exon 11-b(same as primer set 5) | 5'-ACAGCTTCTAATAAAGAGATAAAAC-3' | 5'-GCCGGCATTTATTATTTTTC-3' | 56.4°C | 1 min | Figure 1 B 7 | 850 bp | |
| exon 11-c(same as primer set 6) | 5'-GTTTCTCCTCAAGCAAATACAA-3' | 5'-GATTTTTTACTTTGTCCAAAGATTCC-3' | 60°C | 1 min | Figure 1 B 8 | 873 bp | |
| exon 11-d(same as primer set 7) | 5'-CTGATCCTGCAGCAAAGACC-3' | 5'-GAAAAACCAATGTTTTTTCTCTCTC-3' | 59.2°C | 1 min | Figure 1 B 9 | 908 bp | |
| exon 11-e | 5'-CATTCTAGTGAAGTGTATAATAAATCAG-3' | 5'-ATTCCCCTAAACTATACATAAAAG-3' | 56.4°C | 2 min | Figure 1 B 10 | 1720 bp | |
| exon 12 | 5'-CAATTCTTTAGTTTTAAAAAATGG GC-3 | 5'- GAAAAAGCTTAGAAAAAGAACTTAAAAAATAC-3' | 59.2°C | 1 min | Figure 1 B 11 | 275 bp | |
| exon 13 | 5'- GTAAATGTTTATAATGTGTAATATACAGGC-3' | 5'-CTGTACCTTCCCTACACTATATTAGTAG-3' | 60°C | 1 min | Figure 1 B 12 | 230 bp | |
| exon 14-15 | 5'-CCAAACTTAAATATTTTCTCCTC-3' | 5'-CAGGGATCCCAGTCTATTC-3' | 60°C | 2 min | Figure 1 B 13 | 1213 bp | |
| exon 16 | 5'-GCAGCAAACCCTTGAATGTAG-3' | 5'-GTCAGGTGAACCGTAATGTG-3' | 60°C | 1 min | Figure 1 B 14 | 552 bp | |
| exon 17-8 | 5'- GGTCTTGTACAGTGTAGTGTTTG-3' | 5'-GTTTTTAAGCAATGGAGCATC-3' | 59.2°C | 2 min | Figure 1 B 15 | 1258 bp | |
| exon 19-20 | 5'- CCATCATGTTTAAATTGAAGTCTC-3' | 5'-CAATTACAGAGGTTAAATCAGAAGCC-3' | 59.2°C | 2 min | Figure 1 B 16 | 739 bp | |
| exon 21-24 | 5'-CTCGATATTTGATTCACCAGC-3' | 5'-CAACAGTCCCTTCCTAACCC-3' | 60°C | 2 min | Figure 1 B 17 | 1739 bp | |
| exon 25 | 5'- CAGTATCACTTTTTCTACATTTTG GTC-3' | 5'-CCCAATTTTCACAGAAGCCAC-3' | 59.2°C | 1 min | Figure 1 B 18 | 471 bp | |
| exon 26 | 5'-GGCTTCCATAGATGTTAGATG-3' | 5'-GGACAACTTGGGATCATTTGC-3' | 50.8°C | 1 min | Figure 1 B 19 | 337 bp | |
| exon 27 | 5'- GCTAAATTGCTGATGTTTTCTAC-3' | 5'-CTGCTGAGTCCTCTAATAAGGC-3' | 60°C | 2 min | Figure 1 B 20 | 1437 bp | |
| exon 25 | 5'- CAGTATCACTTTTTCTACATTTTGGTC-3' | 5'-CCCAATTTTCACAGAAGCCAC-3' | 59.2°C | 1 min | Figure 1 B 18 | 471 bp | |
| exon 26 | 5'-GGCTTCCATAGATGTTAGATG-3' | 5'-GGACAACTTGGGATCATTTGC-3' | 50.8°C | 1 min | Figure 1 B 19 | 337 bp | |
| exon 27 | 5'- GCTAAATTGCTGATGTTTTCTAC-3' | 5'-CTGCTGAGTCCTCTAATAAGGC-3' | 60°C | 2 min | Figure 1 B 20 | 1437 bp | |
| For detection of splicing variants | exon 1-11 | 5'-CGAATTTGTTAGCCGTCTCC-3' | 5'-GGATCCTGAGATATTATTTTATTATTAG-3' | 60°C | 2.5 min | Figure 1 C 1 | 2118 bp |
| exon 10-14 | 5'-CTGAAGGGATGTCCAATGC-3' | 5'-GAAATTTGGATTCTGTATTTCTTG-3' | 58°C | 6 min | Figure 1 C 2 | 5594 bp and 1554 bp | |
| exon 11-18 | 5'-CTTCCTGTGAAAACAAATATAG-3' | 5'-GCTGATCTTCTGCTTTTATC-3' | 50.8°C | 2 min | Figure 1 C 3 | 1417 bp | |
| exon 15-25 | 5'-CCTCTGCATGTTCTCATAAAC-3' | 5'-GTGTAAGGTTTAATAATGTCTTCA-3' | 60°C | 2 min | Figure 1 C 4 | 1759 bp | |
| exon 24-27 (same as primer set 13) | 5'-CCTATCCCAAGTTTATCAGCC-3' | 5'-CAGACACAAGTTGATGTTCTCC-3' | 60°C | 2 min | Figure 1 C 5 | 959 bp | |
| For nested PCR of the transcripts lacking most of exon 11 | exon 10-13(1735-7280) | 5'-GTTCTCAAATAATATGACTAATCCAAAC-3' | 5'-GTTCCTCAGTTGTGCGAAAG-3' | 58°C | 6 min | Figure 2 A | 5546 bp and 1506 bp, 1185 bp, 1674 bp or 1270 bp |
| For DNA sequence | cB2 seq1 | 5'-CAATAGAGGTGTTTTCTCCATC-3' | |||||
| cB2 seq2 | 5'-GGATCCTGAGATATTATTTTATTATTAG-3' | 5546 bp and 1506 bp, 1185 bp, 1674 bp or 1270 bp | |||||
| cB2 seq3 | 5'-CCAGCTTTGTCTTTAACCAG-3' | ||||||
| cB2 seq4 | 5'-CTGTGTGACCACTTTCACTATC-3' | ||||||
| cB2 seq5 | 5'-CCCTCCTTCATAAACTGGC-3' | ||||||
| cB2 seq6 | 5'-CTTTCTGAGAGGCATGATCTG-3' | ||||||
| cB2 seq7 | 5'-GCATGGCAAGTGTCTGATTTAC-3' | ||||||
| cB2 seq8 | 5'-GTGAACAAACTTCACAACTTAACC-3' | ||||||
| cB2 seq9 | 5'-GCTGATCTTCTGCTTTTATC-3' | ||||||
| cB2 seq10 | 5'-GGTATGTTTTACAATGATGC-3' | ||||||
| cB2 ex14 R (exon) | 5'-CTAAAGGTTCTTTTTCATTCTTTG-3' | ||||||
| cB2 ex15 F | 5'- GCTTTTTAAATGTTACATGGAGG-3' | ||||||
| cB2 ex17 R | 5'-GTACCAGTCAGGGATGTGAG-3' | ||||||
| cB2 ex18 F (exon) | 5'-ATATGATGTGGAAATTGATAAAA G-3' | ||||||
| cB2 ex22F | 5'-CTTTTTAAAGGGATTCATTTACAG TGG-3' | ||||||
| cB2 ex23 F (exon) | 5'-CCATCACCAGATTTATATTCCC-3' | ||||||
| cB2 ex26 R (exon) | 5'-CAGAAATTTATTTCCTATGCC-3' | ||||||
| cB2 ex23 F (exon) | 5'-CCATCACCAGATTTATATTCCC-3' | ||||||
| cB2 ex26 R (exon) | 5'-CAGAAATTTATTTCCTATGCC-3' | ||||||