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Decontamination of PCR Master Mixes

APPLICATION overview, CHALLENGES AND SOLUTION

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Decontamination of PCR Master Mixes
Decontamination of PCR master mixes
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  • Product overview
  • Performance data & figures
  • Specifications
  • Documents
  • FAQs
  • Ordering Info
  • Protocols
  • Publications

Application Overview

Polymerase chain reaction (PCR) is a highly sensitive technique used in both research and diagnostics for detecting DNA. However, PCR master mixes are often contaminated with E.coli DNA and other elements, which can lead to reduced sensitivity and false positives, especially when targeting small amounts of bacterial DNA. Ensuring the decontamination of PCR master mixes is therefore crucial for the reliability of PCR assays.

See enzymes for

For Detailed Info Including:
  • Product overview
  • Performance data & figures
  • Specifications
  • Documents
  • FAQs
  • Ordering Info
  • Protocols
  • Publications

Application Overview

Polymerase chain reaction (PCR) is a highly sensitive technique used in both research and diagnostics for detecting DNA. However, PCR master mixes are often contaminated with E.coli DNA and other elements, which can lead to reduced sensitivity and false positives, especially when targeting small amounts of bacterial DNA. Ensuring the decontamination of PCR master mixes is therefore crucial for the reliability of PCR assays.

THE PROBLEM These ENZYMEs SOLVE

Decontamination of PCR master mixes

PCR master mixes are susceptible to various sources of contamination, including E.coli DNA, dNTPs, buffer components, and primers/probes. This contamination is particularly problematic when using qPCR for the detection or quantification of minor amounts of bacterial DNA, as it can result in false positives. In assays sensitive to bacterial DNA contamination, a decontamination process is therefore essential for accurate results.

THE PROBLEM These ENZYME SOLVES

Decontamination of PCR master mixes

PCR master mixes are susceptible to various sources of contamination, including E.coli DNA, dNTPs, buffer components, and primers/probes. This contamination is particularly problematic when using qPCR for the detection or quantification of minor amounts of bacterial DNA, as it can result in false positives. In assays sensitive to bacterial DNA contamination, a decontamination process is therefore essential for accurate results.

The presence of E.coli 23S DNA in 2x probe master mixes from various suppliers was quantified by using water instead of template (NTC) and following the manufacturer’s instructions
Fig 1. The presence of E.coli 23S DNA in 2x probe master mixes from various suppliers was quantified by using water instead of template (NTC) and following the manufacturer’s instructions

The presence of E.coli 23S DNA in 2x probe master mixes from various suppliers was quantified by using water instead of template (NTC) and following the manufacturer’s instructions

The figure shows plots from several separate experiments. Traces of E.coli 23S DNA were found in all master mixes tested, with Cq values generally ranging from 30 – 35.

The presence of E.coli 23S DNA in 2x probe master mixes from various suppliers was quantified by using water instead of template (NTC) and following the manufacturer’s instructions

The Solution

Remove contaminating DNA in master mixes without reducing PCR sensitivity
To address this issue, we have developed a PCR Decontamination kit designed to remove contaminating DNA in master mixes without reducing PCR sensitivity. This kit offers a simple and accurate way to ensure that your PCR assays are free from bacterial DNA contamination, thereby enhancing the reliability and accuracy of your results.

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The presence of E.coli 23S DNA in 2x probe master mixes from various suppliers was quantified by using water instead of template (NTC) and following the manufacturer’s instructions
Fig 1.  The presence of E.coli 23S DNA in 2x probe master mixes from various suppliers was quantified by using water instead of template (NTC) and following the manufacturer’s instructions

The figure shows plots from several separate experiments. Traces of E.coli 23S DNA were found in all master mixes tested, with Cq values generally ranging from 30 – 35.

Application Background