Polymorphisms in the Haem Oxygenase-1 promoter are not associated with severity of Plasmodium falciparum malaria in Ghanaian children

Table 1 Primer sequences and conditions for the polymerase chain reactions (PCRs)

Allele specific PCRs	Sequences
Fw	5′-ACTGGCACTCTGCTTTATGTGTGA-3′
Rw A(−413)	5′-GGAGGCAGCGCTGCTCAGAGTAAT-3′
Rw (−413)T	5′-GGAGGCAGCGCTGCTCAGAGTAAA-3′
Conditions	95°C 15 min, 35 cycles: (94°C 30 sec, 60°C 30 sec, 72°C 30 sec), 72° 10 min
Restriction fragment length polymorphism
Fw	5′-TTATTTTATATTTTGTAGAGCC-3′
Rw	5′-AGATGATTCATACAGTCCTTTC-3′
Conditions	94°C 15 min, 45 cycles: (94°C 30 sec, 49°C 30 sec, 72°C 3 min), 72° 10 min
(GT) _n repeat length polymorphism
Fw (5’fam)	5′-AGAGCCTGCAGCTTCTCAGA-3′
Rw	5′-ACAAAGTCTGGCCATAGGAC-3′
Conditions	95°C 15 min, 30 cycles: (95°C 30 sec, 64°C 30 sec, 72°C 30 sec), 72° 10 min

Primer sequences and conditions for the PCR reactions used to determine the polymorphisms in the HMOX1 promoter. The primer pairs for the allele specific PCRs and the restriction fragment length PCR contains a mismatch (Italic). The forward primer for the (GT)_n repeat length PCR is fluorescein-conjugated (highlighted). Primers for the allele specific PCRs were designed based on the HMOX1 nucleotide sequence (Genbank S58267). The primers for the restriction fragment length PCR and (GT)_n repeat length PCR were designed by He et al. [30] and Takeda et al. [33], respectively.

ISSN: 1475-2875