| Gene name | BSP 5' end sequence | BSP 3' end sequence | ΔCt | 3’ end mispriming | Specificity for typical qRT-PCR |
| AXIN1 | GCACCCTCCAAGCAGAG | GAC | 17.6 | yes | yes |
| ACCCTCCAAGCAGAG | GACAA | 20,8* | no | yes | |
| CCCTCCAAGCAGAG | GACAAGA | >17,7** | no | yes | |
| CCCTCCAAGCAGAG | GACAAGAT | 17.6 | yes | yes | |
| CCCTCCAAGCAGAG | GACAAGATC | 11.8 | yes | no | |
| CCAAGCAGAG | GACAAGATCGC | 5.5 | yes | no | |
| CAAGCAGAG | GACAAGATCGCAG | 3.1 | yes | no | |
| BCAS1 | GTCTTTTCTGTGGAGTCTGATGT | CAT | >26,0 | no | yes |
| TCTTTTCTGTGGAGTCTGATGT | CATTT | >27,5 | no | yes | |
| TTTCTGTGGAGTCTGATGT | CATTTGT | >30,0 | no | yes | |
| CTTTTCTGTGGAGTCTGATGT | CATTTGTC | 22.4 | yes | yes | |
| CTGTGGAGTCTGATGT | CATTTGTCT | 18 | yes | yes | |
| GTGGAGTCTGATGT | CATTTGTCTGA | 13.9 | yes | no | |
| TGGAGTCTGATGT | CATTTGTCTGAGA | 11.1 | yes | no | |
| GGAGTCTGATGT | CATTTGTCTGAGAAA | 4.8 | yes | no | |
| BTC | GACGCCCTCCTGTGT | CCC | >16,4 | no | yes |
| CGCCCTCCTGTGT | CCCTC | >21,1 | no | yes | |
| GCCCTCCTGTGT | CCCTCTT | >29,2 | no | yes | |
| GCCCTCCTGTGT | CCCTCTTC | 21.1 | yes | yes | |
| CCCTCCTGTGT | CCCTCTTCG | 12.8 | yes | no | |
| CTCCTGTGT | CCCTCTTCGGA | 8.4 | yes | no | |
| TCCTGTGT | CCCTCTTCGGAAA | 6.9 | yes | no | |
| PCSK6 | CGTAGCTGGCGTAGGAATC | CAG | >32,1 | no | yes |
| GCTGGCGTAGGAATC | CAGGT | >32,1 | no | yes | |
| GCTGGCGTAGGAATC | CAGGTAC | 23.5 | yes | yes | |
| CTGGCGTAGGAATC | CAGGTACC | 18.2 | yes | yes | |
| GGCGTAGGAATC | CAGGTACCA | 13.4 | yes | no | |
| GCGTAGGAATC | CAGGTACCACA | 9.5 | yes | no | |
| CGTAGGAATC | CAGGTACCACATG | 6.0 | yes | no | |
| CGTAGGAATC | CAGGTACCACATGTT | 3.3 | yes | no | |
| * 5' end mispriming observed | |||||
| ** primer-dimers observed | |||||
| Primers to generate the templates (common fixed opposing primer in bold) | |||||
| Gene name | Forward | Reverse | |||
| AXIN1 | ACCCTCCAAGCAGAGGTATGT | TAGCTGCCCTTTTTGGTCAGCA | |||
| BCAS1 | AACAGTGGACCTCAACGAAGGA | AGTCTGATGTTTGGCAGCTGGA | |||
| BTC | TTGGTCATCGGTGTCTGCACAT | ACTTTGCAGCTTGCCACCAA | |||
| PCSK6 | TGAAATGGCTCCAGCAACAGGA | CACGCTGCCTGGACATTCATT | |||
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