Details of Oligonucleotide Primers Used in This Study
| Exon . | Primer Orientation-150 . | External Primer-151 . | Internal Primer . | Fragment Size (bp) . |
|---|---|---|---|---|
| 2 | F | −I1 | A: 5′-GAGAtctagaTTTTGTTTCTGAGCTG-3′ | 891 |
| R | I2 | B: 5′-CAAgaattcAACAGCTTTCTATAG-3′ | ||
| 3 | F | −I2 | b: 5′-ATAAGTGaATTCTCAGTCGTTCTGGTGA-3′ | 309 |
| R | I3b-152 | I3-151 | ||
| 4-153 | F | −I3 | e: 5′-CTCAGGAATTCCACCAAC-3′ | 414 |
| 5-153 | R | I5 | s: 5′-ATTCTGCATGGCGATCGTGG-3′ | 532 |
| 6 | F | −I5 | −I5b: 5′-TTTATAAAATGTTCCTGCAGGATC-3′ | 415 |
| R | q-152 | f-151 | ||
| D-155: 5′-AGTCATCCATCTCAATTTC-3′ |
| Exon . | Primer Orientation-150 . | External Primer-151 . | Internal Primer . | Fragment Size (bp) . |
|---|---|---|---|---|
| 2 | F | −I1 | A: 5′-GAGAtctagaTTTTGTTTCTGAGCTG-3′ | 891 |
| R | I2 | B: 5′-CAAgaattcAACAGCTTTCTATAG-3′ | ||
| 3 | F | −I2 | b: 5′-ATAAGTGaATTCTCAGTCGTTCTGGTGA-3′ | 309 |
| R | I3b-152 | I3-151 | ||
| 4-153 | F | −I3 | e: 5′-CTCAGGAATTCCACCAAC-3′ | 414 |
| 5-153 | R | I5 | s: 5′-ATTCTGCATGGCGATCGTGG-3′ | 532 |
| 6 | F | −I5 | −I5b: 5′-TTTATAAAATGTTCCTGCAGGATC-3′ | 415 |
| R | q-152 | f-151 | ||
| D-155: 5′-AGTCATCCATCTCAATTTC-3′ |
Lowercase indicates non–PIG-A sequence designed to introduce restriction enzyme sites.
F, forward; R, reverse.
Oligonucleotide sequence as reported by Nafa et al.15
New external primers: “I3b”: 5′-gggaaTTCCTATTTATATAAAAATT-3′ and “q”: 5′-AAAATATTGAATGATATAGAGGTAGCATAAC-3′ at position nt 1595-1565 in exon 6.
In the primary PCR, exons 4 + 5 PCR amplified in one fragment by using primers “−I3” and “I5.”15 In the secondary PCR, exons 4 and 5 are individually amplified by using primer “e” (position nt 1014-994 in exon 5) and primer “s” (position nt 938-958 in exon 4).
Specific oligonucleotide that spans the insertion (underlined).