Quantitative Real-Time PCR for Fly Transcripts

Total RNA was extracted from dissected tissues of 10 flies and converted to cDNA using the FastGene Scriptase II cDNA Kit (#LS53, Nippon Genetics Europe Co. Ltd., Duren, Germany). For Real Time-Q-PCR analysis, HOT FIREPol^® EvaGreen^® qPCR Mix Plus (#08-36-00001, Solis BioDyne, Tartu, Estonia) was used. Primers were designed using the primer BLAST tool (http://www.ncbi.nlm.nih.gov/tools/primer-blast, accessed on 17 September 2021) and were as reported before [25 (link),26 (link)]. The ribosomal gene RpL32 (also known as Rp49) was used as an input reference.

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Manola M.S., Gumeni S, & Trougakos I.P. (2021). Differential Dose- and Tissue-Dependent Effects of foxo on Aging, Metabolic and Proteostatic Pathways. Cells, 10(12), 3577.

Publication 2021

FastGene Scriptase II cDNA Kit HOT FIREPol® EvaGreen® qPCR Mix Plus

Cdna Flies Gene Primer Real time pcr analysis Ribosomal Tissues

Corresponding Organization : National and Kapodistrian University of Athens

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Variable analysis

independent variables

Tissue type (dissected tissues of 10 flies)

dependent variables

Gene expression levels measured by Real Time-Q-PCR

control variables

The ribosomal gene RpL32 (also known as Rp49) was used as an input reference

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