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

From: Genetic and biochemical studies in Argentinean patients with variegate porphyria

 

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

  1. 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.