Primers and assay conditions for long and short range PCR reactions
PCR Fragmenta | Primer Sequence 5′ to 3′ | Annealing Temperature | Product Length | Reference Pattern | Variant Pattern |
|---|---|---|---|---|---|
| °C | |||||
| Long PCR on genomic DNA | |||||
| A | 5′ GCCACCATGGTGTCTTTGCTTTCCTGG | 68 | ∼9.5 kb | ||
| 5′ CCGGATTCCAGCTGGGAAATGCG | |||||
| B | 5′ AGGAGGCAAGAAGGAGTGTCAGG | 68 | ∼10.5 kb | ||
| 5′ CCGGATTCCAGCTGGGAAATGCG | |||||
| C | 5′ ATGGCAGCTGCCATACAATCCACCTG | 68 | 6.6 kb | ||
| 5′ ACTGAGCCCTGGGAGGTAGGTAG | |||||
| D | 5′ AGCCACTCTCGTGTCGTCAGCTT | 68 | 3.6 kb (*36) | ||
| 5′ CAGGCATGAGCTAAGGCACCCAGAC | ∼5 kb (*5) | ||||
| Short PCR on genomic DNA | |||||
| E | 5′ AGCCACTCTCGTGTCGTCAGCTT | 68 | 597 bp | absent | 597 bp |
| 5′ CGACTGAGCCCTGGGAGGTAGGTAG | |||||
| 5′ CACCTCTGCTAAGGGAAACAGGCC | 860 bp | 860 bp | 860 bp | ||
| 5′ GCCAGCCTGAACATCCTTTTTGCTA | |||||
| PCR-RFLP on long PCR template | |||||
| F | 5′ CTTCCGTGGAGTCTTGCAGG | 58 | 775 bp | 591 + 184 bp NcoI | 775 bp |
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| 5′ CCTGGGAGGTAGGTAGCCCTG |
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