Table 2. Primers used for the two different multiplex setups.
| Setup 1 | Primers | Concentration [nM] | Origin | Amplicon Tm [°C] | Amplicon Size [bp] | Location [bp] |
| SRY (Control) | GAATATTCCCGCTCTCCGGA/GCTGGTGCTCCATTCTTGAG | 125 | Simoni et al. [10] | 86.50–86.68 | 470 | 2655107–2655576 |
| sY254 (AZFc, DAZ1-4) | GGGTGTTACCAGAAGGCAAA/GAACCGTATCTACCAAAGCAGC | 50 | 81.51–81.83 | 380 | 25316193–25316572; 25372576–25372955; 26952264–26952643; 26986963–26987342 | |
| sY133 (AZFb) | ATTTCTCTGCCCTTCACCAG/TGATGATTGCCTAAAGGGAA | 500 | NEW | 83.86–84.11 | 177 | 23499309–23499485 |
| sY85 (AZFa) | GCTATTCTCTCTCTGGCATC-TGTATT/TGGCAATTTGCCTAT- GAAGT | 375 | NEW | 76.61–76.81 | 80 | 14638102–14638181 |
| Setup 2 | ||||||
| AMELxy (Control) | ATCAGAGCTTAAACTGGG-AAGCTG/CCCTGGGCTCTG-TAAAGAATAGTG | 250 | NEW | 79.79–80.02 | 106 (X) 112 (Y) | 11314994–11315099 (X); 6737888–6737999 (Y) |
| sY255 (AZFc, DAZ1-4) | GTTACAGGATTCGGCGTGAT/CTCGTCATGTGCAGCCAC | 125 | Simoni et al. [10] | 84.35–84.52 | 124 | 25314817–25314940; 25374208–25374331; 26950888–26951011; 26988595–26988718) |
| sY127 (AZFb) | GGCTCACAAACGAAAAGAAA/CTGCAGGCAGTAATAAGGGA | 300 | Simoni et al. [10] | 82.21–82.28 | 274 | 22570417–22570690 |
| G34990 (AZFa) | CATTCGGTTTTATCAGCCAG/CAGTGACTCGAGGTTCAATG | 750 | NEW | 76.41–76.56 | 83 | 15032211–15032293 |
| Alternatively: AMELy (Control) | ATCAGAGCT TAAACTGGG-AAGCTG/CTCTGTAAAGAA- TAGTGGGTGGAT | 500 | NEW | 79.33–79.56 | 105 | 6737888–6737992 |
Given are their sequences, the final concentration in the PCR reaction, the origin (either Simoni et al. [10] or newly designed), the amplicon melting temperatures (given as regions), amplicon size and the Y-chromosomal location based on Human Genome GRCh37/hg19 assembly (Feb. 2009).