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Fig. 1 | Malaria Journal

Fig. 1

From: Bead-based assays to simultaneously detect multiple human inherited blood disorders associated with malaria

Fig. 1

Genomic location and amplification of SNP targets. a Top: Representation of the multi-exon G6PD gene on the X chromosome. The G202A and the A376G markers are amplified by the primer pairs G202A Fw (forward)/G202A Rv (reverse) and A376G Fw (forward) and A376G Rv (reverse). The G202A allele specific primer extension (ASPE) probes (G202 and A202) anneal to the sense strand and either have a G or an A at their 3′ end (see red highlighted letters in brackets) and the A376G probes (A376 and G376) anneal to the sense strand and either have an A or a G at their 3′ end (see red highlighted letters in brackets). Bottom: Representation of the multi-exon HBB gene on chromosome 11. The HbS and HbC markers are amplified by the HBB prime pair. The HbS ASPE probes anneal to the sense strand and the HbC ASPE probes anneal to the anti-sense strand (see red highlighted letters in brackets). b G6PD and HBB amplification. Three primer concentrations were tested (conditions 1–3) on human positives (wells 1–22) and negative (N) controls. Condition 1 = equimolar primer concentration (200 nM each), condition 2 = 100 nM each of G202A and A376G primers and 300 nM each of HBB primers and condition 3 = 150 nM each of G202A and A376G primers and 300 nM each of HBB primers. Even numbered wells correspond to TA = 62.5 °C and odd numbered wells to TA = 60.2 °C. The HBB amplicon is 802 bp long (blue arrow), the G6PD A376G amplicon is 769 bp long (white arrow) and the G6PD G202A amplicon is 619 bp long (yellow arrow)

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