Table 1. Primer and Taqman probe sequences used for multiplex qPCR and optimal ß-actin concentrations.
| Transcript | Sequences | Optimal ß-actin concentration |
|---|---|---|
| ß-actin | Forward: 5′ TGTGCTGCTTACAGAGG 3′ Reverse: 5′ GTACGACCAGAGGCCTA 3′ Probe: 5′/CY5/GCCAACAGAGAAAAGATGACACAGATC 3′ | 500 nM (c)/200 nM (i) 500 nM (c)/200 nM (i) 250 nM (c)/100 nM (i) |
| Dmrt1 canonical | Forward: 5′ CCAACACATTCAACAAACA 3′ Reverse: 5′ ACTGCTGTAGTAGGTGGAGTC 3′ Probe: 5′/FAM/ATCAGAGGGACGGATGCTCATTCAG 3′ | 500 nM 500 nM 300 nM |
| Dmrt1 isoform | Forward: 5′ TACTCCTCGCCACTGAA 3′ Reverse: 5′ CACTCTGGCCCAGGTAG 3′ Probe: 5′/FAM/TGGCAGCCAGATGAAAAGCACAG 3′ | 600 nM 600 nM 300 nM |
Notes.
- (c)
- ß-actin concentration for PCR of Dmrt1 canonical transcript
- (i)
- ß-actin concentration for PCR of non-canonical isoform