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Gene Expression Profiling: qPCR Toolkit for Quality Control
Švec D., Jacobsson J., Sjöback R., Kubista M.

TATAA Biocenter explain how they developed and optimized high-throughput gene expression qPCR with ValidPrime quality control, compensating for inter-run variations.

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Rapid PCR for Integration in Sample-to-answer Analysis Platforms
S. Brunklaus, T.E. Hansen-Hagge, J. Erwes, J. Höth, M. Jung, D. Latta, X. Strobach, C. Winkler, T. Röser, M. Ritzi-Lehnert, K.S. Drese

This poster describes how molecular testing at the point-of-care can increase time to results and yield rather specific information, concentrating on PCR on a chip layout which proves to be fast and robust.

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Hot Start Amplification using OligoBeads via Gradual Release of Bound Primers
Dr. Nam Ngo, Dr. Laurent Jacquinod

OligoBeads provide a mean to store normalized primers used in performing enzymatic reactions including PCR. Primer bound beads eliminate the potential for pipeting errors and reduce contamination thus yielding lower repeat rates and less reagent wastage. The primers bound to the OligoBeads can be stored over a period of a few months without degradation in a nuclease free environment.

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Target length effect on sensitivity and specificity of oligonucleotide microarrays: Advantages of a modified PCR based labelling method over the dendrimer technology.
Abdullah Gibriel1*, Walter Kolch2 and Andrew Pitt1,3

Several methods have been developed for target labelling to enable DNA microarray quantification without taking careful consideration for target length effect. This report highlights the importance of choosing the optimum target length that would ensure specificity without compromising sensitivity of the assay. It also shows the advantages of using the modified PCR method over other methods in generating labelled amplicons of the desired lengths to maximize hybridization efficiency.

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On chip micro-extraction and real-time PCR with integrated SPAD optical fluorescence detection for nucleic acid analysis
Cristina Potrich, Elisa Morganti, Nicola Massari, Lucio Pancher, C. Kostoulas, Laura Pasquardini, Cristian Collini, Andrea Adami, Lorenzo Lunelli, F. Kalatzis, David Stoppa, Cecilia Pederzolli, Leandro Lorenzelli

A PDMS lab-on-a-chip for one step DNA isolation and real time-polymerase chain reaction (RT-PCR) has been designed, fabricated, and characterized for point-of-care clinical diagnostics. In addition, a module for on-chip optical detection based on SPAD - Single-Photon Avalanche Diode - detector has also been developed and used to monitor the presence of specific DNA polymorphisms possibly related to genetic diseases.

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Chemical Variant of 7-deaza-dGTP for Improved CG-rich PCR Amplification
E. H. Ashrafi, S. Shore, T. Le, V. Timoshchuk, N. Paul, R. Hogrefe, G. Zon, I. Koukhareva, A. Lebedev

PCR amplification of nucleic acids is a fundamental technique used in many molecular biology laboratories. Despite its wide use, certain GC-rich regions of DNA, such as mycobacterial disease targets, still remain a challenge for amplification. Sequences high in GC content are associated with the formation of secondary structure, which prevents adequate strand separation and DNA polymerase amplification. As a consequence, mispriming is prominent, complicating specific product formation.

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Periodontal Pockets Microbiota qPCR Analysis at Different Stages of Periodontitis
Oksana Zorina and Denis Rebrikov

Six major periodontal pathogens were quantified (with standardization to human gDNA) in periodontal pockets of periodontitis patients and healthy subjects by original qPCR assay. Relative amount of pathogens to total bacterial mass increased for P.gingivalis (100-fold), P.intermedia (10-fold) and T.forsythensis (10-fold).

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Low-volume on-chip single sperm cell analysis
Pickrahn I. E. ,Schmidt-Gann G., Kroneis T.

We performed low-volume on-chip DNA typing of single sperm cells isolated by means of laser microdissection. 16plex PCR was successful in 39 of 40 single cell samples yielding a mean PCR efficiency of 62.8%. In addition, we were able to identify a single-cell sample containing more than one cell enabling us to monitor the quality of the whole procedure, and hence, exclude “contaminated” samples from further analysis.

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Development and NDA-Level Validation of a Real-Time PCR Procedure for Detection and Quantification of Residual E.coli DNA Contamination of Biopharmaceutical Products
Dan Papa, Pedro J. Morales and Michael D. Sadick

Escherichia coli (E. coli) has been commonly used for the production of biopharmaceuticals. Among the impurities that must be monitored in biopharmaceuticals is residual host-cell DNA (HCD). This study describes the development and subsequent NDA-level validation of a real time PCR procedure developed in response to a client’s need to improve the sensitivity of detection and quantification of residual E. coli HCD in their drug product.

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Scientific News
Lab-Tested Diagnosis Needed When Treating Persistent Diarrhea
New PCR multiplex method makes lab testing more effective.
Biomarker for Multiple Sclerosis Detection Discovered
Winthrop-University Hospital researchers discover biomarker for multiple sclerosis detection.
Scientists Link Bipolar Disorder to Unexpected Brain Region
Researchers from The Scripps Research Institute have found that gene within the brain’s striatum could be linked to biopolar disorder.
Quick, Early Test For Ebola Could Prevent Epidemics
Researchers from Princeton University are collaborating with U.S. government labs to develop a more rapid, accurate and inexpensive test for the Ebola virus with the aim of identifying infections before carriers become symptomatic and contagious.
From Super to Ultra-Resolution Microscopy
A new method pushes the frontier in imaging resolution, with the potential to distinguish individual features in single molecules.
Cancer Research UK joins forces with U.S. 'Cancer Moonshot'
Cancer Research UK and the US government’s National Cancer Institute have announced that two teams will work together to radically accelerate progress against cancer, in one of the first international collaborations inspired by US Vice President Joe Biden’s Cancer Moonshot.
Assessing the Effectiveness of Genome-Editing Technologies
Researchers have developed a cost-effective and rapid method for assessing edits generated by CRISPR-Cas9 and other genome-editing technologies.
Measuring the Abundance of Extremely Rare Mutations
Researchers from Rutgers University demonstrate the use of multiplex real-time PCR assays to measure the abundance of extremely rare mutations associated with cancer.
Harnessing Helpful Microbes
Seeking to further harness microbes’ many uses, the federal government has launched the National Microbiome Initiative (NMI) to “foster the integrated study of microbiomes across different ecosystems.”
What Makes a Good Scientist?
It’s the journey, not just the destination that counts as a scientist when conducting research.

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