Polymerase Chain Reaction (PCR)

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High-throughput PCR based diagnostics: Linking sample handling to molecular oncology risk groups

24 June 2010 | By Ehsan G. Karimiani, Stephan Mohr & Philip J. R. Day, University of Manchester

Cancer molecular pathology broadly relies on the comparison between diseased and normal tissues, with statistically validated differences revealing cancerassociated pathways. This approach, although comparatively one-dimensional, has been remarkably successful, enabling identification of many types of malignant biomarkers and providing the means to develop pharmaceutical agents directed against pertinent biological targets.…

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There is more to qPCR than the PCR reaction

29 September 2008 | By

The polymerase chain reaction is arguably the most significant technical discovery yet to have been made in the field of molecular biology and genetics, if not all life science. It cannot be overstated how much of an impact this technique has had, resulting in molecular biology becoming an integral part…

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Real-time quantitative PCR – opportunities and pitfalls

2 August 2008 | By

The emergence of next generation sequencing technology has brought the prospect of digital analyses closer, technology that will allow not just the quantification of nucleic acids, but will result in the fine-tuning of this information with respect to tissue- and cell-specific transcription, the identification of new transcriptional units, e.g. the…

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Night on Highway 128

19 March 2008 | By

Most people in molecular biology today are not old enough to remember pre-PCR. But try to do your job without it and you will see what a difference that simple little technique has made. ‘Polymerase Chain Reaction’ is now a word in Merriam Webster’s Collegiate Dictionary and if you put…

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The role of chemokines in type 1 diabetes: as assessed by RT-PCR

19 March 2008 | By

Type 1 diabetes is an autoimmune disease, characterised by immune infiltration into the islets of Langerhans, resulting in the destruction of insulin producing b-cells. Over recent years, evidence has been collected on the important role of chemokines in the recruitment of immune cells leading to the pathology of this disease.…

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Analysis of microRNA expression by qPCR

23 November 2007 | By

Alteration of microRNA (miRNA) expression in a disease compared to a healthy state and/or correlation of miRNA expression with clinical parameters (like disease progression or therapy response), may indicate that miRNAs can serve as clinically relevant biomarkers1-3. An important first step for further functional characterisation is the information about differential…

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Cutting edge technologies and their potential role in pharmaceutical microbiology

23 November 2007 | By

In order to meet the challenges demanded by the requirements of Process Analytical Technology (PAT), the modern microbiological laboratory needs to become more innovative in microbial detection, identification and enumeration. Technology is becoming available that will speed up microbiological analysis, potentially allowing pharmaceutical microbiology tests to get as close as…

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The real-time reverse transcription polymerase chain reaction – treat with caution

21 September 2007 | By Stephen A. Bustin, Academic Department of Surgery, Institute of Cell and Molecular Science, Queen Mary’s School of Medicine and Dentistry, University of London

The real-time reverse transcription polymerase chain reaction (RT-qPCR) has become the enabling technology par excellence in every field of molecular research and development, including that of clinical drug development and discovery. Its ability to detect as well as quantify RNA biomarkers sensitively, specifically and speedily has made it an indispensable…

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Real-time PCR gene expression profiling

25 January 2007 | By Mikael Kubista, TATAA Biocenter and MultiD Analyses AB, Sweden, Björn Sjögreen, Center for Applied Scientific Computing, Lawrence Livermore National Laboratory, United States and MultiD Analyses AB, Amin Forootan, MultiD Analyses AB, Radek Sindelka and Jiri Jonák, Laboratory of Gene Expression, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic and José Manuel Andrade, Dept of Analytical Chemistry, University of A Coruna, Spain

Real-time PCR has rapidly become the preferred technique for quantitative analysis of nucleic acids. Its superior sensitivity, reproducibility and dynamic range make it the preferred choice for expression profiling in scientific, as well as routine, applications.

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Quantitative PCR assays in clinical drug development

28 September 2006 | By Michael E. Burczynski, Ph.D. and Ole E. Vesterqvist, Ph.D. Biomarker Laboratory, Clinical Translational Medicine, Wyeth Research

Biomarkers (biological markers) have become an integral part of both drug discovery and drug development and play an important role in the transition of potential new drugs from discovery into clinical drug development. In the past, most biomarkers were proteins/peptides and metabolites measured by technologies such as immunoassays, enzymatic assays,…

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Real-time PCR: clinical applications

24 March 2006 | By Lut Overbergh and Chantal Mathieu, Laboratory for Experimental Medicine and Endocrinology (LEGENDO), University Hospital Gasthuisberg, Catholic University of Leuven

The real-time PCR technique is one of the emerging techniques that, although only described for the first time about a decade ago, have become the method of choice for quantification of DNA and RNA levels in cells, tissues and tissue biopsies.

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Making the lab workhorse run faster

2 February 2006 | By Holly Hilton, Ph.D. and Charu Kanwal Ph.D. Research Informatics, Genetics and Genomics Hoffmann La-Roche Inc.

Quantitative real-time PCR, the workhorse of any genomics lab, is a well established technique that has numerous uses due to its simplicity and flexibility. In this article we will review a brief history of real time PCR, discuss our strategy for optimising a lab running Quantitative-RT-PCR and describe how this…

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Options for quantitative analysis by real-time PCR

11 November 2005 | By Gareth Elvidge, PhD, Genomics Group, Wellcome Trust Centre for Human Genetics, University of Oxford

The expansion of microarray-based gene expression studies has led to an increase in demand for gene-specific PCR-based methods for independent validation of results. Although a number of technologies are available to meet this requirement the most popular is currently real-time PCR.