Total RNA was isolated from ~2,000 synchronized day 5-adult worms using RNAbee (Tel-Test Inc.). cDNA was generated using qScript Flex cDNA synthesis kit (Quantabio). SybrGreen real-time qPCR experiments were performed with a 1:20 dilution of cDNA using a CFC384 Real-Time System (Bio-Rad). Data were analysed with the comparative 2ΔΔCt method using the geometric mean of cdc-42 and pmp-3 as housekeeping genes90 (link). See Supplementary Table 2 for details about the primers used for this assay.
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Quantitative Gene Expression Analysis in C. elegans
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Corresponding Organization : University of Cologne
Other organizations : University of California, Berkeley
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Variable analysis
independent variables
- Not explicitly mentioned
- Gene expression levels
- Total RNA isolated from ~2,000 synchronized day 5-adult worms
- CDNA generated using qScript Flex cDNA synthesis kit
- SybrGreen real-time qPCR experiments performed with a 1:20 dilution of cDNA
- Data analyzed with the comparative 2ΔΔCt method
- Geometric mean of cdc-42 and pmp-3 used as housekeeping genes
- Positive control: None mentioned
- Negative control: None mentioned
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