Pcr Inhibitor Removal
Pcr Inhibitor Removal - Related Videos
01:52
Eukaryotic Transcription Inhibitors
0 Views •
2020
Certain biochemical processes, such as embryonic development and cell growth regulation, depend on the repression of specific genes. DNA binding proteins known as eukaryotic transcription inhibitors regulate the repression of gene expression in eukaryotes. The presence of these inhibitors at the required location and time in the cell is triggered by the presence of hormones and additional signals from other cells. Eukaryotic transcription inhibitors usually contain two distinct domains, a DNA...
08:51
Genotyping of Staphylococcus aureus by Ribosomal Spacer PCR (RS-PCR)
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Cited by 9 •
2016
Here, ribosomal spacer PCR (RS-PCR) is used together with a miniaturized electrophoresis system as a fast and high resolution method for genotyping S. aureus at moderate costs allowing a high throughput.
06:01
Optimized PCR-based Detection of Mycoplasma
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Cited by 14 •
2011
The LookOut Mycoplasma PCR Detection Kit utilizes the polymerase chain reaction (PCR), which is established as the method of choice for highest sensitivity in the detection of Mycoplasma, Acholeplasma, and Ureaplasma contamination in cell cultures and other cell culture derived biologicals.
03:41
Agarose Gel Electrophoresis of DNA Amplicons Post PCR: A Method to Analyze Products of Multiplex PCR and Evaluate PCR Reaction Success
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2025
In this video, we demonstrate the separation of bacterial PCR-amplified DNA using agarose gel electrophoresis. Agarose gel functions as a molecular sieve, enabling the negatively-charged DNA to migrate based upon their size under an applied electric field.
08:45
Assessing Cellular Target Engagement by SHP2 (PTPN11) Phosphatase Inhibitors
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Cited by 4 •
2020
The ability to assess target engagement by candidate inhibitors in intact cells is crucial for drug discovery. This protocol describes a 384 well format cellular thermal shift assay that reliably detects cellular target engagement of inhibitors targeting either wild-type SHP2 or its oncogenic variants.
FAQs
Humic acids, heme, polysaccharides, and residual detergents can reduce PCR performance by binding nucleic acids or DNA polymerase, sequestering essential cofactors, or otherwise disturbing the amplification reaction. Because these contaminants act through different mechanisms, a sample may show reduced efficiency, sensitivity, specificity, or reproducibility rather than a single predictable failure pattern.
Removing inhibitors restores access to the molecular components required for amplification. If a contaminant binds DNA polymerase, associates with nucleic acids, or sequesters cofactors, the reaction cannot proceed with its expected efficiency. Cleanup therefore affects not only whether a product appears, but also the assay’s sensitivity, specificity, and reproducibility.
Controlled dilution lowers the concentration of inhibitory substances in the extract before PCR. Its value lies in reducing interference enough to permit amplification while retaining sufficient template for detection. Dilution is therefore one option within PCR inhibitor removal, alongside washing, adsorption, precipitation, and filtration, rather than a universal replacement for those approaches.
PCR inhibitor removal may use washing, adsorption, precipitation, filtration, or controlled dilution, depending on which contaminants remain in the extract. These operations reduce inhibitor concentration or separate interfering substances from nucleic acids. The practical goal is a cleaner template that supports more dependable downstream PCR amplification.
Sample context influences the cleanup challenge because inhibitor classes can occur in biological, soil, food, and environmental extracts. The presence of humic acids, heme, polysaccharides, or residual detergents means that extract quality cannot be judged by the source label alone. Cleanup is therefore part of preparing complex templates for reliable PCR analysis.
Effective cleanup supports accurate pathogen detection, genotyping, forensic analysis, and molecular ecology. By improving amplification sensitivity, specificity, and reproducibility, it helps researchers interpret nucleic acid measurements from complex samples with greater confidence. This is especially important when biological, soil, food, or environmental extracts contain substances that could otherwise compromise PCR results.