Table 1 Overview of the genotyped SNVs with the primer and probe sequences, amplicon lengths and annealing temperatures (Ta) of the qPCR assays. Target SNVs are indicated in bold, interfering SNVs are underlined

1

GB54921- RA:r.94G > A

F1: 5′-ACCCACTTTTTACTACGA-3′

R1: 5′-GCTTCTAGGCTGGATAA-3’

Wt1-probe: 5’-FAM-TGGACAAATTTACCTTCTCGTTA-BHQ1–3′

Vt1-probe: 5′-TexasRed- TGGACAAATTTATCTTCTCGT-BHQ2–3’

108 bp

58 °C

2

GB54921- RA:r.144 A > G

F1: 5‘-CCAAGTTCCCGTCAGA-3’

R1: 5′-TCGCCATTCTTCTCAGG-3’

Wt1-probe: 5’-FAM-CTCTAACGATGCTTCTAGGC-BHQ1–3′

Vt1-probe: 5’-TexasRed-CTCTAACGACGCTTCTAGGC-BHQ2–3’

106 bp

58 °C

3

GB47018- RA:r.1824C > U

F1: 5′-AAGGGACTAACTATAGCAAAA-3′

R1: 5′-GGCAGGAGGTGTTTTAG-3’

Wt1-probe: 5’-FAM-CGAATCGCTCCCGGAAA-BHQ1–3′

Vt1-probe: 5′-TexasRed-CGAATCGCTTCCGGAA-BHQ2–3’

90 bp

60 °C

4

GB53345- RA:r.37 A > GG

F1: 5’-AGCGATAAAATTTCTTCTTTCCTTA-3′

F2: 5′-AGCGATAAAATTTCTTCTTTCTTTATC-3′

R1: 5’-CATCGTCCTGGCGTAG-3’

Wt1-probe: 5’-FAM-AGCGTCATCGCCGTC-BHQ1–3’

Vt1-probe: 5’-TexasRed-AGCGTCACCGCCGTC-BHQ2–3’

Vt2-probe: 5’-TexasRed-CAGTGTCACCGCCGTC-BHQ2–3’

118 bp

58 °C

5

GB53340- RA:r.4143 U > G

F1: 5′-CGAAGGTGGCCGAATTG-3′

R1: 5′-GCTTCTCCAACTCGTTCATC-3’

Wt1-probe: 5’-FAM-TCGGGAGGTTCTCATCCACC-BHQ1–3′

Vt1-probe: 5′-TexasRed-AATCGGGAGGTGCTCATCCA-BHQ2–3′

Vt2-probe: 5′-TexasRed-AATCGGGAGGTGCTGATCCA-BHQ2–3’

147 bp

60 °C

6

GB48382- RA:r.987 G > A

F1: 5‘-TGGCGAATGGGAAACAG-3’

R1: 5′-CTCGTACCTTTTCAGTCTTCA-3’

Wt1-probe: 5’-FAM-CGTTTATACGCGCCATTTTTCGA-BHQ1–3′

Vt1-probe: 5′-TexasRed-CGTTTATACGTGCCATTTTTCG-BHQ2–3’

132 bp

62 °C

7

GB50526- RA:r.1662G > A

F1: 5‘-CGTGATCGTCGGTGTTATC-3’

R1: 5′-GCGAGAGGGTGAAGGA-3’

Wt1-probe: 5’-FAM-TCTCCTTTCGGGTCGGCTG-BHQ1–3′

Vt1-probe: 5′-TexasRed-TCTCCTTTTGGGTCGGCT-BHQ2–3’

84 bp

62 °C

8

GB50114- RA:r.1662A > G

F1: 5‘-CTCTGAACACCCTGAACAAG-3’

R1: 5′-TCCAGCTCCTGTCCTTG-3’

Wt1-probe: 5’-FAM-TACTGCCCCTGGTGGC-BHQ1–3′

Vt1-probe: 5′-TexasRed-TTACTGCCCCCGGTGGC-BHQ2–3’

138 bp

62 °C