Steady-State Transcript Levels of the Porphobilinogen Deaminase Gene in Patients with Acute Intermittent Porphyria

Table 4.

Oligonucleotides Used in the RT, PCR, and SPMS Reactions

Mutation Primer sequence 5‘ → 3‘ Direction Size of PCR product (bp) Incorporated [3H]dNTPs in SPMS reaction (normal/ mutant allele)
517C → T  1. PCR: ATGATGGCACTGAACTCCT antisense
 and  2. PCR: Bio-CGGGGAAACCTCAACAC sense 62
 518G → A  3. MS/517: GTCCAGCTTCCGAAGCC antisense GGG/A
 4. MS/518: GTCCAGCTTCCGAAGC antisense C/T
673C → G  5. RT and PCR:  GCGATGCAGCGAAGCAGAGTC antisense G/CC
 and  6. PCR: Bio-CGGCCTTGGGCGTGGAAG sense 94  and
 673C → T  7. MS: GATGTCCTGGTCCTTGGCTC antisense  G/A
713T → G  8. PCR: CCTTTCAGCGATGCAGCGAAGC antisense
 9. PCR: = 6. sense 101 A/C
10. MS: AGCAGAGTCTCGGGATCGTGC antisense
1073delA 11. PCR: CATGAAGATGGCCCTGAGGAT sense
12. PCR: Bio-GGCATCTGTGCCCCACAAACCAG antisense 197 A/C
13. MS: GGATGTTGCACGGCAGCTTA sense

  • ↵Specific activities for tritium-labeled nucleotides: [3H]dATP, 59 Ci/mmoles; [3H]dCTP, 64 Ci/mmoles; [3H]dGTP, 31 Ci/mmoles; [3H]dTTP; 119 Ci/mmoles. (Bio) 5‘biotinylated primer; (MS) solid-phase minisequencing detection primer.

This Article

  1. Genome Res. 7: 1054-1060

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