Table 4. .
Primers and Restriction Enzymes Used for Confirmation of RAB23 Mutations[Note]
| Mutation and Primer | Primer Sequence (5′→3′)a | Product Size (bp) |
Restriction Enzyme |
| E48fsX7: | 157 | XcmI (−) | |
| E48fsXdigF | AAAGACTACAAGAAAACCATTGCCATTG | ||
| RAB23_2R | AGTTGCCACACCTCGAAATC | ||
| Y78fsX30: | 188 | StuI (−) | |
| RAB23_3F | TTACCAAAAACATTTTCCTTTACA | ||
| RAB23_3R | GCCAAAATAATATGCCCAAA | ||
| C85R: | 156 | BssSI (+) | |
| C85RdigF | TTTGAATGGATAAAAGTTGCCC | ||
| C85RdigR | TTCCCTATCTGTGGTAGAGAACTCGAGC | ||
| E137X: | 207 | HindIII (+) | |
| RAB23_5F | AAACAAGCTATCAGAAGGCACC | ||
| RAB23_5R | CAACACAATTTTAAAAGCGCA | ||
| L145X: | 120 | HpaI (−) | |
| RAB23_5F | AAACAAGCTATCAGAAGGCACC | ||
| RAB23_L145XdigR | TTCTTTCACTGATGTTCTGTAGAATGTT |
Note.— PCRs were performed using the same conditions as described in table 1.
a
Nucleotides shown in bold represent mismatches incorporated into primers to engineer diagnostic restriction sites.