A simple method for testing and controlling inhibition in soil and sediment samples for qPCR

Tom Bott; George Shaw; Simon Gregory

doi:10.1016/j.mimet.2023.106795

A simple method for testing and controlling inhibition in soil and sediment samples for qPCR

J Microbiol Methods. 2023 Sep:212:106795. doi: 10.1016/j.mimet.2023.106795. Epub 2023 Aug 3.

Authors

Tom Bott¹, George Shaw², Simon Gregory³

Affiliations

¹ British Geological Survey, Keyworth, Nottingham NG12 5GG, UK; School of Biosciences, Sutton Bonington, University of Nottingham, Loughborough LE12 5RD, UK. Electronic address: tom.bott@nottingham.ac.uk.
² School of Biosciences, Sutton Bonington, University of Nottingham, Loughborough LE12 5RD, UK.
³ British Geological Survey, Keyworth, Nottingham NG12 5GG, UK. Electronic address: simongr@bgs.ac.uk.

PMID: 37541445
DOI: 10.1016/j.mimet.2023.106795

Abstract

The presence of polymerase-chain-reaction (PCR) inhibitors in many environmental samples can make reliable and repeatable quantitative-polymerase-chain-reaction (qPCR) analysis difficult without sample dilution. To estimate an optimal sample dilution for qPCR and reduce effects of inhibition, a simple test based on multiple dilution series of samples is presented that avoids the use of internal controls and standards reducing complexity and cost.

Keywords: Inhibition; Method optimisation; Quantitative-PCR (qPCR).

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Polymerase Chain Reaction / methods
Real-Time Polymerase Chain Reaction
Soil*

Substances

Soil

Grants and funding

BB_/Biotechnology and Biological Sciences Research Council/United Kingdom