|Novel, Fully Automated Method Allows Efficient Analysis of qPCR Data for Qualitative Calling Based on Comparative Cq|
Collaborative research by Pioneer Hi-Bred (a DuPont company) and Azure PCR Limited
Assessment to ascertain if a method for analysis of qPCR data dependent on manual intervention can be replaced by automated analysis using the AzurePCRTM method.
|Volume-Related Inhibitors Standardization for Reverse Transcription Quantitative Polymerase Chain Reaction Experiments|
Pascal Pugniere, Sebastien Banzet, Thomas Chaillou, Catherine Mouret and Endre Peinnequin
This poster addresses the reliability of qPCR data and its dependence on technical variations. The proposal is that constant volume of RNA extract can improve reliability of RT-qPCR.
|Defining off-target cleavage in a pair of Zinc Finger Nucleases|
K. Mukherjee, D. Carroll
This study looks at off-target cleavage of Zinc Finger Nucleases (ZNFs) in Drosophila in an attempt to analyze potential cleavage spots, with a view to designing more efficient ZFNs.
| 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.
|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.
|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.
|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.
|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.
|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.