| Primer name | Sequence (5′-3′) | Nested PCR reaction (product size) | Diagnostic for |
| To test F8 exon 22-23 integrity and detect inverted F8 exon 22 to intron 22 mRNA product | |||
| Exon 22 FWD1 | GTGGATCTGTTGGCACCAATG | First PCR | Exon 22-23 junction |
| Exon 24 REV | CTCCCTTGGAGGTGAAGTCG | (428 bp) | |
| Exon 22 FWD2 | ACCAATGATTATTCACGGCATCAAGA | Second PCR | Exon 22-23 junction |
| Exon 23 REV | TGCAAACGGATGTATCGAGCAATAA | (225 bp) | |
| Exon 19 FWD1 | TCCAAAGCTGGAATTTGGCG | First PCR | Intron 22 inverted RNA |
| INT22 REV | CAATTCTTTCCATTTTCCAAGACACCGTG | (428 bp) | |
| Exon 19 FWD2 | TGCTGGGATGAGCACACTTTT | Second PCR | Intron 22 inverted RNA |
| INT22 REV | CAATTCTTTCCATTTTCCAAGACACCGTG | (378 bp) | |
| To test F8 Exon1-2 integrity and detect inverted F8 exon1-VBP1 chimeric mRNA product | |||
| SP FWD | GCACATCCAGTGGGTAAAGTTC | First PCR | Exon 1-2 junction |
| Exon 3 REV | AGACTGACAGGATGGGAAGC | (509 bp) | |
| SP FWD | GCACATCCAGTGGGTAAAGTTC | Second PCR | Exon 1-2 junction |
| Exon 2 REV | GGCCTTGGCTTAGCGATGTT | (413 bp) | |
| Exon 1 FWD1 | TGGGAGCTAAAGATATTTTAGAGAA | First PCR | F8-VBP1 chimeric RNA |
| VBP1 REV1 | ACTTATACTTCTGGTACTGTTCATCCAGC | (590 bp) | |
| Exon 1 FWD2 | GAATTAACCTTTTGCTTCTCCAGTTGAAC | Second PCR | F8-VBP1 chimeric RNA |
| VBP1 REV2 | TGTTTCATGAAGGAATCTAC | (500 bp) |
| Primer name | Sequence (5′-3′) | Nested PCR reaction (product size) | Diagnostic for |
| To test F8 exon 22-23 integrity and detect inverted F8 exon 22 to intron 22 mRNA product | |||
| Exon 22 FWD1 | GTGGATCTGTTGGCACCAATG | First PCR | Exon 22-23 junction |
| Exon 24 REV | CTCCCTTGGAGGTGAAGTCG | (428 bp) | |
| Exon 22 FWD2 | ACCAATGATTATTCACGGCATCAAGA | Second PCR | Exon 22-23 junction |
| Exon 23 REV | TGCAAACGGATGTATCGAGCAATAA | (225 bp) | |
| Exon 19 FWD1 | TCCAAAGCTGGAATTTGGCG | First PCR | Intron 22 inverted RNA |
| INT22 REV | CAATTCTTTCCATTTTCCAAGACACCGTG | (428 bp) | |
| Exon 19 FWD2 | TGCTGGGATGAGCACACTTTT | Second PCR | Intron 22 inverted RNA |
| INT22 REV | CAATTCTTTCCATTTTCCAAGACACCGTG | (378 bp) | |
| To test F8 Exon1-2 integrity and detect inverted F8 exon1-VBP1 chimeric mRNA product | |||
| SP FWD | GCACATCCAGTGGGTAAAGTTC | First PCR | Exon 1-2 junction |
| Exon 3 REV | AGACTGACAGGATGGGAAGC | (509 bp) | |
| SP FWD | GCACATCCAGTGGGTAAAGTTC | Second PCR | Exon 1-2 junction |
| Exon 2 REV | GGCCTTGGCTTAGCGATGTT | (413 bp) | |
| Exon 1 FWD1 | TGGGAGCTAAAGATATTTTAGAGAA | First PCR | F8-VBP1 chimeric RNA |
| VBP1 REV1 | ACTTATACTTCTGGTACTGTTCATCCAGC | (590 bp) | |
| Exon 1 FWD2 | GAATTAACCTTTTGCTTCTCCAGTTGAAC | Second PCR | F8-VBP1 chimeric RNA |
| VBP1 REV2 | TGTTTCATGAAGGAATCTAC | (500 bp) |
The protocol for both the reverse transcription PCR and the nested PCR steps of the Inv22 assay was as follows: 50°C, 30 min; 95°C, 15 min; 30 cycles of 94°C (30 s), 50°C (45 s), and 72°C (60 s) followed by a 10-min incubation at 72°C. The protocol for both the reverse transcription PCR and the nested PCR steps of the Inv1 assay was as follows: 50°C, 30 min; 95°C, 15 min; 30 cycles of 94°C (30 s), 59°C (45 s), and 72°C (60 s) followed by a 10-min incubation at 72°C.