For each time point/treatment,four biological replicates, consisting of four individuals each, were
analysed. All reactions were prepared as triplicates and a no-template
control was included for each primer pair. The expression of
three putative reference genes was examined and two with stable
expression in the investigated samples were selected for normalization:
actin and cyclophilin. The PCR program consisted of: 95 C
for 5 min, 45 cycles of 95 C for 10 s, 58 C for 20 s and 72 C for
30 s. A melting curve was performed starting at 95 C for 5 s,
65 C for 5 min and continuously increasing by 0, 11 C/s until the
temperature reached 97 C. To calculate the relative transcript ratio
between a sample and a control, a formula by Pfaffl
For each time point/treatment,four biological replicates, consisting of four individuals each, wereanalysed. All reactions were prepared as triplicates and a no-templatecontrol was included for each primer pair. The expression ofthree putative reference genes was examined and two with stableexpression in the investigated samples were selected for normalization:actin and cyclophilin. The PCR program consisted of: 95 Cfor 5 min, 45 cycles of 95 C for 10 s, 58 C for 20 s and 72 C for30 s. A melting curve was performed starting at 95 C for 5 s,65 C for 5 min and continuously increasing by 0, 11 C/s until thetemperature reached 97 C. To calculate the relative transcript ratiobetween a sample and a control, a formula by Pfaffl
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