Primers and conditions for PCR amplification of NAG exons
| Exon | Primer sequence | Annealing temp (°C) | MgCl2 (mmol/l)/ DMSO) (%) | Product size (bp) | Restriction enzyme before SSCP (fragment sizes (bp)) |
|---|---|---|---|---|---|
| 1 | F: AGACGCCCCCAAGGGAGTAT | 57 | 1.5/10 | 608 | BssHII (315, 235, 58) |
| R: ATTTGGGTGGCAGCGGCTCC | |||||
| 2 | F: CCCTGCCCATCTGTTAGACT | 55 | 1.5/10 | 596 | AvaII (308, 203, 85) |
| R: GCACGTTGAAAGCACTTCTA | |||||
| 3–4 | F: AGCGCCCAGCACAAAGAAG | 57 | 1.0 | 561 | PstI (315, 246) |
| R: AAATTCCCTCTCTGAGCTAA | |||||
| 5 | F: AAACCAGGAGCTGTAGAGAAGT | 54 | 1.5 | 434 | BstXI (259, 175) |
| R: CTGCCTACCCCTACTGACATCT | |||||
| 6.1 | F: GGCCCTCTGTTTCATCACTC | 57 | 1.0 | 444 | SphI (251, 193) |
| R: AAATCTGGCACTGGGTCCTT | |||||
| 6.2 | F: GCATCAGCCAGAACGAAGTG | 52 | 1.5 | 436 | PstI (236, 190) |
| R: CCAGCTCCTTGCTCAGGTAG | |||||
| 6.3 | F: CAACCGATCTGATGTGTTTG | 55 | 1.5 | 387 | HhaI (234, 153) |
| R: TTGGCATAGTCCAGGATGTT | |||||
| 6.4 | F: GCCGAGGCCGATTTCTAC | 57 | 1.0 | 345 | SfaNI (180, 165) |
| R: GCGAATCTATCACCAAGAGC | |||||