Expected molecular weights (bp) of amplicons from IS-PCR and RT-NPCR assays to detect intron 22 inversions
| F8 allele . | IS-PCR diagnostic Inv22 . | IS-PCR complementary Inv22 . | First RT-NPCR . | Second RT-NPCR . |
|---|---|---|---|---|
| Male | ||||
| N | 487 | 457; 405 | 225 | — |
| Inv22-1 | 333 | 559; 457 | — | 378 |
| Inv22-2 | 385 | 559; 405 | — | 378 |
| Inv22-3* | NR | NR | — | 378 |
| Dup 1-22* | 487 | 559; 457; 405 | 225 | 378 |
| Del 22-1* | 333 | 457 | — | — |
| Del 22-2* | 385 | 405 | — | — |
| Female (or male with >1 F8 gene or Xq28 segment) | ||||
| N/N | 487 | 457; 405 | 225 | — |
| N/Inv22-1 | 487; 333 | 559; 457; 405 | 225 | 378 |
| N/Inv22-2 | 487; 385 | 559; 457; 405 | 225 | 378 |
| N/Inv22-3* | NR | NR | 225 | 378 |
| N/Dup 1-22* | 487 | 559; 457; 405 | 225 | 378 |
| N/Del 22-1* | 487; 333 | 457; 405 | 225 | — |
| N/Del 22-2* | 487; 385 | 457; 405 | 225 | — |
| F8 allele . | IS-PCR diagnostic Inv22 . | IS-PCR complementary Inv22 . | First RT-NPCR . | Second RT-NPCR . |
|---|---|---|---|---|
| Male | ||||
| N | 487 | 457; 405 | 225 | — |
| Inv22-1 | 333 | 559; 457 | — | 378 |
| Inv22-2 | 385 | 559; 405 | — | 378 |
| Inv22-3* | NR | NR | — | 378 |
| Dup 1-22* | 487 | 559; 457; 405 | 225 | 378 |
| Del 22-1* | 333 | 457 | — | — |
| Del 22-2* | 385 | 405 | — | — |
| Female (or male with >1 F8 gene or Xq28 segment) | ||||
| N/N | 487 | 457; 405 | 225 | — |
| N/Inv22-1 | 487; 333 | 559; 457; 405 | 225 | 378 |
| N/Inv22-2 | 487; 385 | 559; 457; 405 | 225 | 378 |
| N/Inv22-3* | NR | NR | 225 | 378 |
| N/Dup 1-22* | 487 | 559; 457; 405 | 225 | 378 |
| N/Del 22-1* | 487; 333 | 457; 405 | 225 | — |
| N/Del 22-2* | 487; 385 | 457; 405 | 225 | — |
Del22-1 and Del 22-2, F8 exon 1-22 deletion involving 0.5 Mb from int22h-1 and the most centromeric of the homologs int22h-2, int22h-3. Dup 1-22, tandem duplication(s) of an F8 gene segment spanning int22h-1 to the most centromeric copy of int22h (int22h-2 or int22h-3). Inv1, intron 1 inversion; Inv22, intron 22 inversion; Inv22-3, type 3 inversions11,12 involve homologous recombination of int22h-1 with a duplicated int22h sequence (a rare genetic event, <1% of F8 inversion mutations); N, normal F8 gene; NR, not reported here, as this family of rare mutations would produce amplicons of various lengths.
Additional F8 gene mutations/rearrangements resulting from alternative pairings among the int22h homologs int22h-1, int22h-2, and int22h-3.14,24