Sample degradation leads to false-positive copy number variation calls in multiplex real-time polymerase chain reaction assays

2009 ◽  
Vol 386 (2) ◽  
pp. 288-290 ◽  
Author(s):  
Holly N. Cukier ◽  
Margaret A. Pericak-Vance ◽  
John R. Gilbert ◽  
Dale J. Hedges
Keyword(s):  
Polymerase Chain Reaction ◽  
Copy Number Variation ◽  
Real Time ◽  
False Positive ◽  
Copy Number ◽  
Chain Reaction ◽  
Number Variation ◽  
Polymerase Chain

Rapid Determination of Perv Copy Number From Porcine Genomic DNA by Real-Time Polymerase Chain Reaction

2011 ◽  
Vol 22 (4) ◽  
pp. 175-180 ◽  
Author(s):  
Donghee Lee ◽  
Jungeun Lee ◽  
Jong-Kwang Yoon ◽  
Na Young Kim ◽  
Gye-Woong Kim ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Copy Number ◽  
Genomic Dna ◽  
Rapid Determination ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Evaluation method for asymmetric uncertainty of quantitative polymerase chain reaction measurements of deoxyribonucleic acids with low copy number

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Unoh Ki ◽  
Takeru Suzuki ◽  
Satoshi Nakazawa ◽  
Yuuki Yonekawa ◽  
Kazuki Watanabe ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Confidence Interval ◽  
Real Time ◽  
Real Time Pcr ◽  
Copy Number ◽  
Limit Of Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Low Copy Number ◽  
Polymerase Chain

AbstractRecently, in food safety and various other fields, qualitative and quantitative gene analysis using real-time polymerase chain reaction (PCR) method has become increasingly popular. The limit of detection (LOD) and quantifiable range for these measurements depends on the range and precision of DNA calibrators’ concentrations. Low-copy-number nucleic acid reference materials with low uncertainty produced by an inkjet system have been developed to allow for precise measurements in a low-copy-number region. However, when using a calibrator with a low copy number near one, the copy number distribution is asymmetric. Consequently, the confidence intervals of estimated copy numbers can include negative values when conventional methods of uncertainty estimation are used. A negative confidence interval is irrelevant in the context of copy number, which is always positive value or zero. Here, we propose a method to evaluate the uncertainty of real-time PCR measurements with representative values and an asymmetric 95% confidence interval. Moreover, we use the proposed method for the actual calculation of uncertainty of real-time PCR measurement results for low-copy-number DNA samples and demonstrate that the proposed method can evaluate the precision of real-time PCR measurements more appropriately in a low-copy-number region.

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