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Accurate Real-time Reverse Transcription Quantitative PCR
2009Within the last few years real-time quantitative PCR has become the method of choice for the accurate quantification of mRNA levels. Compared to previous methods the sensitivity of real-time quantitative PCR improved to the detection limit of up to one single molecule per reaction tube.
Marco, Klatte, Petra, Bauer
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High‐Throughput Real‐Time Quantitative Reverse Transcription PCR
Current Protocols in Molecular Biology, 2006AbstractExtensive detail on the application of the real‐time quantitative polymerase chain reaction (QPCR) for the analysis of gene expression is provided in this unit. The protocols are designed for high‐throughput, 384‐well‐format instruments, such as the Applied Biosystems 7900HT, but may be modified to suit any real‐time PCR instrument. QPCR primer
Angie L, Bookout +4 more
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Quantitative Real-Time PCR in aDNA Research
2011Quantitative real-time PCR (qPCR) is a technique that is widely used in the field of ancient DNA (aDNA). Quantitative PCR can be used to optimize aDNA extraction methodologies, to detect PCR inhibition, and to quantify aDNA libraries for use in high-throughput sequencing.
Bunce, Michael, Oskam, C., Allentoft, M.
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New FRET primers for quantitative real-time PCR
Analytical and Bioanalytical Chemistry, 2007FRET primer real-time PCR chemistry depends on internally labeled primers with FRET dyes linked to their 3' end. The best distance between the FRET dyes for obtaining the largest signal and the lowest background is six nucleotides. In this study the forward primer was labeled with FAM and the reverse primer with Texas red; the labeled primers meet in ...
Ashraf I, Ahmad, Jahan B, Ghasemi
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Evaluation of uncertainty in quantitative real-time PCR
Journal of Microbiological Methods, 2006Quantitative real-time PCR is one of the newer methods for measurement of the amount of nucleic material in biological systems. However, reliable measurement requires an appropriate estimation of uncertainty and this paper has developed the uncertainty budget associated with this procedure using as an example, data from a quantitative real-time PCR ...
John L, Love +5 more
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Quantitative detection of periodontopathogens by real-time PCR
Journal of Microbiological Methods, 2004Specific bacteria are believed to play an important role in chronic periodontitis, yet the significance of their relative numbers in initiation and progress of the disease is still unclear. We report here the development of a sensitive, quantitative PCR technique for enumerating Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg ...
Claudia, Nonnenmacher +3 more
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Real-Time Quantitative PCR, Pathogen Detection and MIQE
2012Nucleic acids are the ultimate biomarker and real-time PCR (qPCR) is firmly established as the method of choice for nucleic acid detection. Together, they allow the accurate, sensitive and specific identification of pathogens, and the use of qPCR has become routine in diagnostic laboratories. The reliability of qPCR-based assays relies on a combination
Gemma, Johnson +2 more
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Quantitative measurement of streptomycetes using real-time PCR
Journal of Environmental Monitoring, 2006A quantitative real-time PCR method was developed and used for determination of streptomycetes in indoor dust samples of five homes collected during three years. The specificity of the method was tested with 14 Streptomyces and ten non-streptomycetous species, revealing a high specificity for mesophilic streptomycetes. Thermophilic species and S. albus
Helena, Rintala, Aino, Nevalainen
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RNA Isolation and Real-Time Quantitative RT-PCR
2008Adipose tissue has emerged as a major endocrine organ producing a wide spectrum of hormones and factors that play crucial roles in regulating cell turnover and function, not only locally within the adipose tissue but also in the brain and other key metabolic organ systems.
Haiyan, Guan, Kaiping, Yang
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Semi-Quantitative Real-Time PCR for Pain Research
2004In this chapter we cover what we have found to be a "best practice" for real-time polymerase chain reaction (PCR) for relative mRNA quantification. We describe our techniques for tissue-sample collection and freezing, sample handling for quick and reproducible extraction of total RNA, first strand cDNA synthesis, real-time PCR amplification, and ...
Hong Qing, Guo, Sandra R, Chaplan
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