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A variety of PCR inhibitors inherent to clinical and environmental samples can negatively impact the sensitivity and accuracy of a molecular assay. Conventionally, to overcome this, specimens undergo processing before testing. However, extraction technologies are not 100% efficient and can impact the amount of target nucleic acid available for testing. Some PCR inhibitors can also co-elute following purification, reducing the efficiency, particularly in quantitative PCR multiplex assays and may even induce false-negative assay results.
Reaction buffer, enhancers and stabilizers, dNTPs and MgCl2, designed to be used in PCR reactions for highly reproducible, accurate assay results in the presence of inhibitors.
Meridian’s universal Inhibitor-Tolerant qPCR Buffer is designed for developing qPCR multiplex assays that require minimal sample processing and fast turn-around times (TAT). Because the buffer is highly resistant to various qPCR inhibitors, it can be used with crude lysates or inhibitor-rich samples, to directly detect pathogens, including viruses and bacteria, in various biological specimens and to analyze gene expression in cells.
| Description | Reaction buffer, enhancers and stabilizers, dNTPs and MgCl2, designed to be used in PCR reactions for highly reproducible, accurate assay results in the presence of inhibitors, such as human and animal blood. |
| Concentration | 5 x |
| Appearance | Clear, colorless solution |
| Application | Inhibitor tolerant PCR |
| Sample type | cDNA, crude or purified DNA |
| Presentation | 1 vial |
| Storage | -20 °C |
| Mix stability | See outer label |
| Consistency | ±0.5 Ct variance between test and reference sample |
| DNA Contamination | None detected in PCR amplification with traces overlay with the negative control on E. coli and mouse genomic DNA specific targets. |
| DNase Contamination | No detectable degradation |
We have tested whole human and bovine blood, saliva, sputum, urine, cerebrospinal fluid (CSF), cow milk, stool sample and plant lysate, other types of samples can be used, but we would recommend testing different concentrations to determine the concentrations that can be tolerated.
Yes, Inhibitor-Tolerant Buffer, 5x requires the addition of DNA polymerase, dNTPs, MgCl2, primers, probes and clinical samples.
The Inhibitor-Tolerant qPCR Buffer can be dried down, but it will require the addition of excipients and glycerol-free polymerase.
We would recommend using a range of crude sample concentrations or extract volumes to ascertain at what point unacceptable PCR inhibition starts to take place. This will vary as some sample types, such as blood and stool material typically have more inhibition.
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