Genetic and biochemical studies in Argentinean patients with variegate porphyria

BMC Medical Genetics

Table 2 Primers and conditions used for PPOX gene and cDNA amplification

	Amplified region (exon)	Amplified Region (intron)	MW (pb)	Sequence (5' to 3')	Annealing (°C)	Mg⁺² (mM)
I	2, 3, 4	2, 3	758	F: GCTTCTGGAGCGCAGGTTGTCC R: AAGGCATATGAGGATGAGGGCA	60	1.6
II	5, 6	5	680	F: AGGTATGTCAGGAGCTTCCCCC R: CCCTCACTTTGGCAGTACTTAA	60	1.6
III	7, 8	7	852	F: TGCTGGGATTACAGGTGT R: AGCTTTTGCTTCTCACTGGTAGG	62	2.5
IV	9	-----------	321	F: GATTACAGGTGTGAGCCACCA R: CCTACCAGTGAGAAGCAAAAGCT	60	2
V	10, 11,12, 13	10,11,12 13	859	F: GCCCTTTCCTTCTGACGCATG R: GCCAGACCAAGCCAAGCCAAGC	62	2.5
VI	1	1	440	F: CCAAGTCCCGCCAATCCAGAT R: GGACAACCTGCGCTCCAGAAGC	60	5
VII	Promotor	-----------	705	F: AGGTGATAGAGAACTGGCCCAA R: CAGCCTTTTCGGTCTCTCCTA	63	3
VIII	------------	4	786	F: TCTGAGCTTGGCTTGGATTC R: CTCTGCAGCTGGTTTAGG	60	2
IX	------------	6	528	F: GCTTTCCCAGTCTCTTCC R: AAGGCCTGGCGAATGAGT	60	2
X	------------	8	313	F: ATTCTCATTTTCTGGGTCTCTC R: TCAGCAGGGAGCAGCTCACTG	60	2
XI	------------	9	535	F: CTGAGTGCCATCACTGCA R: GCTCAGGGAAAGCAACTG	60	2
XII	Gen	All	5,500	F: AGAGAACTGGCCCAAAATTGGAGT R: CAGACCAAGCCAAGCCAAGCAATT	60	2
4–9	From nt 226 (exon 4) to nt 893 (exon 9).		668	F: TCTGAGCTTGGCTTGGATTC R: GTCACTCGACGAGGGACGACT	60	3
6–9	From nt 478 (exon 6) to nt 893 (exon 9).		416	F: TCTCTAGCCATGGACAGTCT R: GTCACTCGACGAGGGACGACT	60	6

All primers were developed in the course of this study. Primers I to XII were used for DNA amplification and primers 4–9 and 6–9 were used for cDNA amplification.

ISSN: 1471-2350