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Table 4.

Oligonucleotides Used in the RT, PCR, and SPMS Reactions

Mutation	Primer sequence 5‘ → 3‘[i]	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

[i] 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.