Development and validation of a high throughput, closed tube method for the determination of haemoglobin alpha gene (HBA1 and HBA2) numbers by gene ratio assay copy enumeration-PCR (GRACE-PCR)

Table 1 Primers used for the GRACE-PCR alpha globin copy number assay

Primer Direction	Primer Sequence 5′-3′	Target Gene symbol	Primer Concentration (μM)	Product size (bp)	Product Tm (°C)	Position on Ref Sequence NC_000016.10
Forward	CACCCGGCCTCATGGAT	CLCN7	0.16	155	79.4	1,462,101 to 1,462,255
Reverse	AAGAGAACTACAGACCAACACCC	CLCN7	0.16	155	79.4	1,462,101 to 1,462,255
Forward	CCATCTTTACGTTTCTGGGCACTC	HBA1	0.45	131	82.2	177,800 to 177,930
Reverse	GCCATGCTGGAGTGGGACTTC	HBA1	0.45	131	82.2	177,800 to 177,930
Forward	CCGTTAAGCTGGAGCCTCGGT*A	HBA2	0.45	171	85.2	173,594 to 173,764
Reverse	ACACCTCCATTGTTGGCACAT	HBA2	0.45	171	85.2	173,594 to 173,764

ISSN: 1471-2350