Investigation on NTC (No-Template Control) Amplification in real time PCR test of Boar DNA sample
Asrori, M. R.; Telaumbanua, G. N. C.; Khoirunnisa, S.; Wonorahardjo, S.; Suharti, S.
Show abstract
Amplification in the No-Template Control (NTC) is a critical issue in real-time PCR (qPCR) as it can compromise the validity of results. This study was conducted to investigate the source of an observed signal in the NTC of a qPCR assay designed for boar DNA detection. Methodologies included melt curve analysis to characterize the amplification product and standard curve analysis to assess assay performance. The results from the melt curve analysis definitively confirmed that the product amplified in the NTC was not a primer-dimer, indicating that the signal did not originate from non-specific primer interactions. Furthermore, the primer set clearly demonstrated efficient amplification of the target DNA sequence in positive samples. The overall assay performance was validated by a standard curve, which showed acceptable linearity with a coefficient of determination (R2) of 1. Collectively, these findings suggest that the qPCR assay is robust and specific. The amplification observed in the NTC is attributed to minute DNA contamination rather than methodological flaws like primer-dimer formation, confirming the high sensitivity of the developed test.
Matching journals
The top 8 journals account for 50% of the predicted probability mass.
Similar papers in this journal
- Validation of a Novel Molecular Assay to the Diagnostic of COVID-19 Based on Real Time PCR with High Resolution Melting 94%
- Quantitative PCR assays to detect humpback whale (Megaptera novaeangliae), shortbelly rockfish (Sebastes jordani), and common murre (Uria aalge) in marine water samples 94%
- Multiplex detection of "Candidatus Liberibacter asiaticus" and Spiroplasma citri by qPCR and droplet digital PCR 94%
Similar papers in this journal
- PCR procedures to amplify GC-rich DNA sequences of Mycobacterium bovis 95%
- Development and Validation of a multi-target TaqMan qPCR method for detection of Borrelia burgdorferi sensu lato 92%
- Microliter spotting and micro-colony observation: a rapid and simple approach for counting bacterial colony-forming units 91%
Similar papers in this journal
- Development and validation of cost-effective one-step multiplex RT-PCR assay for detecting the SARS-CoV-2 infection using SYBR Green melting curve analysis 94%
- A Paper-based Loop-Mediated Isothermal Amplification (LAMP) Assay for Highly Pathogenic Avian Influenza 94%
- A simple RNA preparation method for SARS-CoV-2 detection by RT-qPCR 94%
Similar papers in this journal
- A broad-based probe-free qPCR assay for detection and discrimination of three human herpes viruses 95%
- Evaluation Of SYBR Green Real Time PCR For Detecting SARS-CoV-2 From Clinical Samples 93%
- Development of a SYBR Green quantitative PCR assay for detection of Lates calcarifer herpesvirus (LCHV) in farmed barramundi 93%
Similar papers in this journal
- Improved and Simplified Diagnosis of Covid-19 using TE Extraction from Dry Swabs 92%
- LAMPrey: a standardised method for analysing quantitative LAMP (qLAMP) and qPCR reactions using the inflection cycle threshold iCt 91%
- MINTyper: An outbreak-detection method for accurate and rapid SNP typing of clonal clusters with noisy long reads 90%
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.