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Updated: Oct 4, 2025

Simultaneous DNA-RNA Extraction from Coastal Sediments and Quantification of 16S rRNA Genes and Transcripts by Real-time PCR
Published on: June 11, 2016
Accurate qPCR quantification in polymicrobial communities requires assessment of gDNA extraction efficiency
Nuno Cerca1, Ângela Lima1, Angela França1
1Laboratory of Research in Biofilms Rosário Oliveira (LIBRO), Centre of Biological Engineering (CEB), University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal; LABBELS -Associate Laboratory, Braga, Guimarães, Portugal.
Abstract:
qPCR absolute quantification requires the assessment of PCR reaction efficiency. This is achieved by performing serial dilutions of gDNA. Herein, we demonstrate that when quantifying bacterial load in mixed samples, reaction efficiency should not be used as a calibration curve since gDNA isolation efficiency is neglected, significantly impacting quantification.
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