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Multi-template polymerase chain reaction
Publication date: Available online 4 December 2014 Source:Biomolecular Detection and Quantification Author(s): Elena Kalle , Mikael Kubista , Christopher Rensing PCR is a formidable and potent technology that serves as an indispensable tool in wide range of biological disciplines. However, due to the ease of use and often lack of rigorous standards many PCR applications can lead to highly variable, inaccurate, and ultimately meaningless results. Thus, rigorous method validation must precede its broad adoption to any new application. Multi-template samples possess particular features, which make their PCR analysis pr...
Source: Biomolecular Detection and Quantification - December 4, 2014 Category: Molecular Biology Source Type: research

Making standards for quantitative real-time pneumococcal pcr
Publication date: Available online 4 December 2014 Source:Biomolecular Detection and Quantification Author(s): Susan C. Morpeth , Jim F. Huggett , David R. Murdoch , J. Anthony G. Scott Quantitative lytA PCR is often performed using in-house standards. We hypothesised equivalence when measuring a standard suspension of Streptococcus pneumoniae by colony-forming-units (CFU) or genome-copies. Median (IQR) ratio of CFU/genome-copies was 0.19 (0.1-1.2). Genome-copies were less variable than CFU, but the discrepancy between the methods highlights challenges with absolute quantification. (Source: Biomolecular Detection and Quantification)
Source: Biomolecular Detection and Quantification - December 4, 2014 Category: Molecular Biology Source Type: research

A novel approach for evaluating the performance of real time quantitative loop-mediated isothermal amplification-based methods
This study uses a sexually transmitted infection diagnostic model in combination with an adapted metric termed isothermal doubling time (IDT), akin to PCR efficiency, to compare quantitative PCR and quantitative loop-mediated isothermal amplification (qLAMP) assays, and to quantify the impact of matrix interference. The performance metric described here facilitates the comparison of qLAMP assays that could assist assay development and validation activities. (Source: Biomolecular Detection and Quantification)
Source: Biomolecular Detection and Quantification - November 22, 2014 Category: Molecular Biology Source Type: research

A survey of tools for the analysis of quantitative PCR (qPCR) data
We report an overview of the functionality, features and specific requirements of the individual software tools, such as data exchange formats, availability of a graphical user interface, included procedures for graphical data presentation, and offered statistical methods. In addition, we provide an overview about quantification strategies, and report various applications of qPCR. Our comprehensive survey showed that most tools use their own file format and only a fraction of the currently existing tools support the standardized data exchange format RDML. To allow a more streamlined and comparable analysis of qPCR data, mo...
Source: Biomolecular Detection and Quantification - November 20, 2014 Category: Molecular Biology Source Type: research

Determining lower limits of detection of digital PCR assays for cancer-related gene mutations
Publication date: September 2014 Source:Biomolecular Detection and Quantification, Volume 1, Issue 1 Author(s): Coren A. Milbury , Qun Zhong , Jesse Lin , Miguel Williams , Jeff Olson , Darren R. Link , Brian Hutchison Digital PCR offers very high sensitivity compared to many other technologies for processing molecular detection assays. Herein, a process is outlined for determining the lower limit of detection (LoD) of two droplet-based digital PCR assays for point mutations of the epidermal growth factor receptor (EGFR) gene. Hydrolysis probe mutation-detection assays for EGFR p.L858R and p.T790M mutations were...
Source: Biomolecular Detection and Quantification - November 20, 2014 Category: Molecular Biology Source Type: research

A current overview of commercially available nucleic acid diagnostics approaches to detect and identify human gastroenteritis pathogens
Publication date: September 2014 Source:Biomolecular Detection and Quantification, Volume 1, Issue 1 Author(s): Kate Reddington , Nina Tuite , Elizabeth Minogue , Thomas Barry Purpose of review Gastroenteritis is caused by a wide range of viral, bacterial and parasitic pathogens and causes millions of deaths worldwide each year, particularly in infant populations in developing countries. Traditional microbiological culture and immunological based tests are time consuming, laborious and often lack diagnostic specificity and sensitivity. As a result patients can receive suboptimal and/or inappropriate antimicrobial t...
Source: Biomolecular Detection and Quantification - November 20, 2014 Category: Molecular Biology Source Type: research

Digital PCR: A brief history
Publication date: September 2014 Source:Biomolecular Detection and Quantification, Volume 1, Issue 1 Author(s): Alexander A. Morley Digital PCR for quantification of a target of interest has been independently developed several times, being described in 1990 and 1991 using the term “limiting dilution PCR” and in 1999 using the term “digital PCR”. It came into use in the decade following its first development but its use was cut short by the description of real-time PCR in 1996. However digital PCR has now had a renaissance due to the recent development of new instruments and chemistry which have made it a much...
Source: Biomolecular Detection and Quantification - November 20, 2014 Category: Molecular Biology Source Type: research

How to make Mathematics Biology's next and better microscope
Publication date: September 2014 Source:Biomolecular Detection and Quantification, Volume 1, Issue 1 Author(s): Jim Huggett , Justin O’Grady , Stephen Bustin (Source: Biomolecular Detection and Quantification)
Source: Biomolecular Detection and Quantification - November 20, 2014 Category: Molecular Biology Source Type: research