Primers (Supplementary Table 1) were designed and synthesised as described previously (Siemienowicz et al., 2020) (link). Quantitative RT-PCR was performed on 384-well plate format (Applied Biosystems) with all samples assayed in duplicate and housekeeping control genes included in each run, as well as template, RNA and RT-negative controls, using the ABI 7900HT Fast Real Time PCR system (Applied Biosystems) as described previously (Hogg et al., 2012) (link). The transcript abundance of target gene relative to the housekeeping genes was quantified using the Ct method (Livak and Schmittgen, 2001) (link).
Genorm reference gene selection kit
The GeNorm Reference Gene Selection Kit is a laboratory tool designed to identify the most stable reference genes for gene expression normalization in qPCR experiments. The kit provides a systematic approach to assess and select the optimal reference genes for a given experimental setup.
Lab products found in correlation
6 protocols using genorm reference gene selection kit
Adipose and Liver RNA Extraction
Primers (Supplementary Table 1) were designed and synthesised as described previously (Siemienowicz et al., 2020) (link). Quantitative RT-PCR was performed on 384-well plate format (Applied Biosystems) with all samples assayed in duplicate and housekeeping control genes included in each run, as well as template, RNA and RT-negative controls, using the ABI 7900HT Fast Real Time PCR system (Applied Biosystems) as described previously (Hogg et al., 2012) (link). The transcript abundance of target gene relative to the housekeeping genes was quantified using the Ct method (Livak and Schmittgen, 2001) (link).
Standardized qRT-PCR Gene Expression
Oxidative Stress Assay in BEAS-2B Cells
Real-time qPCR analysis of mRNA expression
Validating Microarray Assays with qPCR
Immunofluorescence Staining of Muscle Cells
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