Data were analyzed using PRISM software version 9.0 (GraphPad Software, Inc., San Diego, CA). Determination of relative gene expression was done in duplicates and represented in graphs plotting the mean ± SD. Our limit of detection (LoD) was calculated using an average of each species probing for Pan Eukaryotic 18S (ThermoFisher Scientific Cat# 4333760F, MA USA) with a cycle threshold of 35. Please note that each sample was subtracted each own 18S value and hence can appear below the LoD but its expression was detected in a Ct value below 35. Samples that did not amplify were given an arbitrary value of 39.99. Variance between the relative expression of genes between organs and by species was determined using one‐way ANOVA. Post hoc analysis was done to determine the difference between the expression of these genes when there was statistical significance determined by one‐way ANOVA.
Pan eukaryotic 18s
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Pan Eukaryotic 18S is a laboratory product designed to detect the 18S ribosomal RNA (rRNA) gene, which is a common genetic marker used to identify eukaryotic organisms. The product provides a standardized method for the detection and analysis of eukaryotic organisms in various samples.
Automatically generated - may contain errors
4 protocols using pan eukaryotic 18s
1
Relative Gene Expression Analysis
2
Quantifying Relative Gene Expression via RT-qPCR
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Relative genetic expression was determined using real‐time quantitative polymerase chain reaction (RT‐qPCR) (CFX96™ Real‐Time System, Bio‐Rad, CA, USA) using commercially available TaqMan primers (Tables 1 , 2 , 3 ) and TaqMan™ Fast Universal PCR Master Mix (2X) no AMPERASE™ UNG (ThermoFisher Scientific, Catalog# #4367846, MA, USA). Amplification was done in 40 cycles with the following conditions (Denaturing at 95°C for 20 seconds and annealing and extending at 60°C for 20 s). Cycle threshold values were normalized using Pan Eukaryotic 18S (ThermoFisher Scientific Cat# 4333760F, MA, USA), transformed using the 2−∆CT method, and graphed to represent the relative genetic expression by sample, gene group, and species.
3
Relative Gene Expression Analysis via RT-qPCR
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Relative genetic expression was determined using Real Time Quantitative Polymerase Chain Reaction (RT-qPCR) (CFX96™ Real-Time System, Bio-Rad, CA, USA) using commercially available TaqMan primers (Tables 1 –3 ) and TaqMan™ Fast Universal PCR Master Mix (2X) no AMPERASE™ UNG (ThermoFisher Scientific, Catalog#4367846, MA, USA). Amplification was done in 40 cycles with the following conditions (Denaturing at 95°C for 20 seconds and annealing and extending at 60°C for 20 seconds). Cycle threshold values were normalized using Pan Eukaryotic 18S (ThermoFisher Scientific, MA, USA), transformed using the 2−∆CT method, and graphed to represent the relative genetic expression by sample, gene group and species.
4
Relative Gene Expression Analysis Across Species
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Data were analyzed using PRISM software version 9.0 (GraphPad Software, Inc., San Diego, CA). Determination of relative gene expression was done in duplicates and represented in graphs plotting the mean±SEM. Our limit of detection (LoD) was calculated using an average of each species probing for Pan Eukaryotic 18S (ThermoFisher Scientific, MA USA) with a cycle threshold of 35. Please note that each sample was subtracted each own 18S value and hence can appear below the LoD but its expression was detected in a Ct value below 35. Samples that did not amplify were given an arbitrary value of 39.99. Variance between the relative expression of genes between organs and by species was determined using one-way ANOVA. Post hoc analysis was done to determine the difference between the expression of these genes when there was statistical significance determined by one-way ANOVA. (*p≦0.05, **p≦0.01, ***p≦0.001 & ****p≦0.0001)
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