Table 1. Sequences of the primers and probes with their respective fluorophore reporter, quencher and nucleotide modifications used in the qPCR screening assay.
| Target | Oligos | Sequence (5’-3’) |
|---|---|---|
| TREC | Fwd primer | TTTGTAAAGGTGCCCACTCCT |
| Rev primer | GCCAGCTGCAGGGTTTAGG | |
| TREC probe | FAM- CGGTGATGC/ZEN/ATAGGCACCTGCACC -3IABkFQ | |
| SMN | Fwd primer | AATGCTTTTTAACATCCATATAAAGCTATC |
| Rev primer | GAATGTGAGCACCTTCCTTC | |
| SMN1 probe* | SUN- AGGG+TT+T+C+AGA+CAA -3IABkFQ | |
| SMN2 blocker* | AGGG+TT+T+T+AGA+CAA | |
| HBB: c.20A>T | Fwd primer | CAACTGTGTTCACTAGCAACC |
| Rev primer | CCCCACAGGGCAGTAACG | |
| HbS allele probe* | Cy5.5- CT+C+CT+G+T+GGAG -3IAbRQSp | |
| HBB blocker* | CT+C+CT+G+A+GGAG | |
| ACTB | Fwd primer | ATTTCCCTCTCAGGCATGGA |
| Rev primer | CACGTCACACTTCATGATGGA | |
| ACTB probe | Cy5- CCTGTGGCA/TAO/TCCACGAAACTACCTTC -3IAbRQSp |
*Bases with + denote locked nucleic acid (LNA™) modifications. Underlined bases indicate nucleotide change of interest for SMN1 exon 7 (c.840C) and HBB: c.20A>T alleles.