Table 3 Primers to amplify target fragments of Dmrt1

Primer name	Primer sequence (5′-3′)	Tm (℃)	Product length
Cyn_Z_8564889_PF	CAACTAAGACTTTTCAAACCC	62	643
Cyn_Z_8564889_PR	TTAAAATCCCAACACTCACAC	62
Dmrt1_exon1_PF	CCATTAATTACAAAATGTTACAGG	60	494
Dmrt1_exon1_PR	AGTAGCAGTAGCAGTATCAGT	60
Dmrt1_exon2_PF	TAAGGCAATAAAGAAAGGTAAGGTC	60	298
Dmrt1_exon2_PR	GCTGTGTTGTGAAGTCTGGT	60
Dmrt1_exon3_PF	TGTCCTTCTATCACTGCATTCTCA	62	276
Dmrt1_exon3_PR	GTGTTTTCCAAATGTATCAGTATGC	62
Dmrt1_exon4_PF	AACAGGTAACAAGTACATTTCTGTG	62	391
Dmrt1_exon4_PR	TGAATGTCGAACGAGCAGGG	62
Dmrt1_exon5_PF	GGAGCGAGTCATTTGATCAGG	60	535
Dmrt1_exon5_PR	TGATTGGATCAGAGCATGATTGT	60

PCR amplification was in a volume of 40 μl, containing 20 μl PCR Mix (TaKaRa), 50 ng template, and 0.5 μM of each primer. The PCR conditions were: 95 °C for 3 min, followed by 32 cycles of 94 °C for 30 s, corresponding melting temperature (Tm) for 30 s, 72 °C for 1 min, and a final extension step at 72 °C for 5 min

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