Table 1.
Primer | Sequence 5′- 3′ | GC | Tm | Size | Eff | R2 |
---|---|---|---|---|---|---|
S18-F |
CAATTAAGGGTGTGGGGCGAAG* |
54.5 |
62.1 |
141 |
99.0 |
1.000 |
S18-R |
TCTTGTATTGGCGTGGATTCTGC* |
47.8 |
60.6 |
|||
SDHA-F |
TGTTTCCCACCAGGTCACACAC* |
54.5 |
62.1 |
119 |
93.4 |
0.991 |
SDHA-R |
CCAGTCGGAGCCCTTCACG* |
68.4 |
63.1 |
|||
PGK1-F |
ACAATGGAGCCAAGTCAG* |
50.0 |
53.7 |
146 |
91.9 |
0.998 |
PGK1-R |
CACGCAGTCCTTCAAGAAC* |
52.6 |
56.7 |
|||
RPL4-F |
CAGACCTTAGCAGAATCTTGAAAAGC* |
42.3 |
61.6 |
171 |
92.0 |
0.998 |
RPL4-R |
CCTGGCGAAGAATGGTGTTCC* |
57.1 |
61.8 |
|||
HSP70-F |
GTCAAGCACGGTGTTCTGTG |
55.0 |
59.4 |
141 |
101.2 |
0.999 |
HSP70-R |
CACGGCAAAGTAGAGATCATCG |
50.0 |
60.3 |
|||
P53-F |
CTCACCATCATCACACTGGA |
50.0 |
57.3 |
175 |
94.2 |
0.998 |
P53-R |
TAGGCAGTGCTCGCTTAGC |
57.9 |
58.8 |
|||
KIN-F |
TGCTGGCTTCAGAAAATCC |
47.4 |
54.5 |
98 | 92.3 | 0.997 |
KIN-R | CTCTTGGTTCCAAAGCGTCTC | 52.4 | 59.8 |
F = forward; R = reverse; GC = GC content (percentage); Tm corresponds to the theoretical melting temperature. R2 corresponds to the linear correlation coefficient of the standard curve obtained by plotting the logarithm of the quantity of gene expression versus the threshold cycle (Ct). The slope of the curve was used to calculate the amplification efficiency (in%) for each pair of primers (Eff = (101/-slope-1)*100). * Primers tested previously in dolphins [13].