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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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Chemically Modified Primers for PCR and Ligation Applications
Elena Hidalgo Ashrafi, Sabrina Shore, Tony Le, Victor Timoshchuk, Natasha Paul, Richard Hogrefe, Inna Koukhareva, Alexandre Lebedev

PCR is an essential tool with utility in a variety of advanced applications. To improve the specificity of PCR, a unique approach to "Hot Start" PCR employing primers containing thermolabile modifications has been developed. These modified primers, named CleanAmp™ Primers, are amenable for use in Hot Start activation schemes as the modification is released after an initial denaturation step.

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Development of a Test Battery for Epigenetic Non-genotoxic Carcinogens
Haroon Rashid

The main objective of this sudy is to obtain an increased insight into the mechanisms of action of epigenetic carcinogens. Although the expected number of non-genotoxic carcinogens among newly registered compounds is unknown, there is a growing concern that when numbers of 2-year cancer bioassays are significantly reduced, non-genotoxic carcinogens may go undetected. Therefore there is a need for the development of alternative methods for their detection.

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Chemically Modified Primers for Improved Multiplex PCR
Elena Hidalgo Ashrafi, Tony Le, Alexandre Lebedev, Richard Hogrefe, Victor Timoshchuk, Sabrina Shore, Inna Koukhareva and Natasha Paul

Multiplex PCR is an advantageous technique used in PCR applications to amplify multiple targets in a single reaction. As useful as it is, this technique presents a new set of challenges that further complicates PCR setup. For example, reactions are more prone to off-target amplifications such as mis-priming and primer dimer due to the increased number of primer pairs.

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Hot Start dNTPs - Novel Chemistries for Use in Advanced PCR Applications
Tony Le, Elena Hidalgo Ashrafi, Sabrina Shore, Victor Timoshchuk, Natasha Paul, Richard Hogrefe, Inna Koukhareva, Alexandre Lebedev

PCR is a widely used scientific tool whose specificity can be increased by the use of Hot Start technologies. Although many Hot Start technologies exist, recently developed CleanAmp™ dNTPs are a distinct approach that employs modified nucleoside triphosphates with a thermolabile protecting group at the 3´-hydroxyl.

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ROLE OF IRF-8 IN THE INTERFACE OF MELANOMA TUMOR CELL-IMMUNE SYSTEM INTERACTION.
Fabrizio Mattei, Giovanna Schiavoni, Massimo Spada, Francesca Spadaro and Lucia Gabriele

Interferon-regulatory factor-8 is essential for differentiation and function of dendritic cells and thus for the induction of competent immune responses. Here, we investigated the role of IRF-8 in affecting immune response against melanoma. Our results reveal a critical role of IRF-8 in controlling melanoma growth and suggest that IRF-8-mediated antitumor activity is the result of a coordinated action between DC-mediated immune response and the tumor suppressor function of IRF-8.

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Scientific News
Understanding Tumor Evolution
Study provides insight into tumor evolution; may point to improved diagnosis and treatment.
Frankfurter Fraud: Finding Out What’s In Your Hot Dog
Scientists have developed a technique to test the meat content of Frankfurters.
How Cloud Connectivity Can Combat the Reproducibility Crisis
This infographic explains the reproducibility crisis, and how cloud connectivity can help overcome this problem.
Keeping Cells in Shape to Fight Sepsis
Boosting levels of a protein that controls the shape and activity of a crucial group of white blood cells improves survival and recovery chances during sepsis.
This 3D Printer Can Detect Infectious Diseases
Researchers at AI Biosciences, Inc. have repurposed 3D printers into machines capable of performing sample preparation and DNA amplification.
Natera, UCSF Announce Collaboration
Joint research program to study DNA markers of kidney transplant rejection.
CDC Updates Zika Recommendations
CDC has issued updated Zika recommendations and guidance for healthcare providers with a focus on sexual transmission.
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.
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