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Changes in the Gene Expression of mRNA Transcripts for Insulin like Growth Factor, their receptor and Facilitative Glucose Transporter in IVM Oocytes
S.C. Gupta, Neelam Gupta and Alok Pandey

The cDNA libraries from single oocytes and pre-implantation buffalo embryos from 2 cell stage to blastocyst were established. Relative expression studied with RT-PCR of IGF-I showed an increase at 12h and decline at 24h of IVM oocytes. IGF-IR was expressed at cleavage stages to blastocyts. Glut-I was expressed in IVM oocytes and SCNT embryos at all stages. Gene expression of IGF-I, IGF-IR and Glut-I plays an important role in SCNT embryo production.

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DNA Methylation Analysis – Reliable Cell Characterization in Regenerative Medicine
Uli Hoffmueller, Stephen Rapko, Udo Baron, Georg Wieczorek, Alexander Hellwag, Cornelia Krüger, Stefan Kärst, Leslie Wolfe and Sven Olek

We demonstrate that DNA methylation patterns can serve as characteristic markers to distinguish different cell types. We have identified panels of methylation markers that are specific to mesenchymal stem cells or various differentiated cell types in the mesenchymal lineage. This method of cell type identification has a number of advantages over conventional markers in that it is robust, is both qualitative and quantitative.

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EM Algorithm for Gene Copy Number Estimation Using TaqMan® Assays
Catalin Barbacioru, Kelly Li, Caifu Chen and Raymond Samaha

Recently, TaqMan® assays have been developed for detection of genetic variation at gene level using primers and probes designed for genomic DNA sequences. The R package TaqGCN contains classes and methods that can be used for data reading and plotting, and for predicting gene copy number.

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EasyBeacons™ - new Probes Ideal for Realtime PCR Detection of Methylation Status of Single CpG Duplets and SNPs
K. Skadhauge, C. Nielsen & U.B. Christensen

The EasyBeacons™ presented here are based on the novel technology Intercalating Nucleic Acid, INA®, linked to a fluorophore and a quencher. INA® is composed of normal DNA nucleotides and Intercalating Pseudo Nucleotides (IPNs). The fact that the EasyBeacons™ are mostly composed of normal DNA nucleotides means that in many respects EasyBeacons™ behave like DNA based probes, allowing use of standard buffers, primers and enzymes and hence reduces the optimisation efforts.

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Real-time PCR Gene Expression Profiling
Mikael Kubista, José Manuel Andrade, Björn Sjögreen and Amin Forootan

Correct interpretation of real-time PCR data requires appropriate experimental design, accurate data pre-processing and analysis of the data using proper statistical and multivariate methods. For this process we have developed the GenEx software.

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Sensitive and Specific Detection of microRNAs
Martin Kreutz, James Qin, Holger Engel, Po-Jen Shih, Peter Hahn, Martin Schlumpberger, Subrahmanyam Yerramilli and Eric Lader

MicroRNAs are endogenous, 21–22 nt, noncoding RNAs that mediate post transcriptional gene regulation. miRNAs are involved in regulation of gene expression during development, differentiation, cell proliferation, and apoptosis. Misregulation of miRNA expression is associated with several cancers and other diseases. We have developed the miScript System for real-time PCR analysis of hundreds of miRNAs, sno RNAs, piRNAs, other small noncoding RNAs.

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Real-time PCR in Diagnostics of Plant Viruses
Mehle N., Boben J. and Ravnikar M.

Sensitive and specific method real-time PCR was developed for diagnostics of certain plant viruses. Real-time PCR can be used for determination of different viruses as well as for differentiation between related strains of viruses. The method can also be used for detection of low concentrations of plant viruses in various samples such as environmental waters, growth substrates and seeds.

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Real-Time Multiplex Rt-PCR on Circulating Tumor Cells
Anieta M. Sieuwerts, Stefan Sleijfer, Jaco Kraan, Joan Bolt-de Vries, Jan-Willem Gratama, John WM Martens and John A. Foekens

Using the CTC kit (CellSearch™), cells that attached to anti-EpCAM Moab were immunomagnetically separated and used for analysis of a selected pilot set of 32 genes by real time RT-PCR. This study shows the feasibility of multiple gene expression analysis on RNA isolated from only one tumor cell. Most importantly, expression analysis of several tumor-specific genes in blood samples containing only 2 tumor cells is already possible.

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Novel Fluidics Microbead Trap/Flow Cell Enhances Speed/Sensitivity of Bead-Based Bioassays Up to 5-Fold
RM Ozanich, CJ Bruckner-Lea, JW Grate, MG Warner, BP Dockendorff, KC Antolick, HC Edberg, LH Johnson, AN Easterday

Pacific Northwest National Laboratory (PNNL) has developed a micro/nano particle trap that allows surface-functionalized magnetic or non-magnetic particles to be trapped with subsequent perfusion of sample, reagents and wash solutions, yielding significant (up to 5-fold) improvements in assay speed and sensitivity, while significantly reducing sample matrix effects.

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Scientific News
The Genetic Roots of Adolescent Scoliosis
Scientists at the RIKEN Center for Integrative Medical Sciences in collaboration with Keio University in Japan have discovered a gene that is linked to susceptibility of Scoliosis.
Diagnostic Test Developed for Enterovirus D68
researchers at Washington University School of Medicine in St. Louis have developed a diagnostic test to quickly detect enterovirus D68 (EV-D68), a respiratory virus that caused unusually severe illness in children last year.
Simple Technology Makes CRISPR Gene Editing Cheaper
University of California, Berkeley, researchers have discovered a much cheaper and easier way to target a hot new gene editing tool, CRISPR-Cas9, to cut or label DNA.
RNAi Screening Trends
Understand current trends and learn which application areas are expected to gain in popularity over the next few years.
HPV Genomes Show Greater Diversity Than Expected in Cancer Patients
The findings could have implications for eventually understanding why some cervical lesions become malignant.
Rapidly Detecting Drug-Resistant HepC
A nested PCR-based assay has been shown to rapidly and accurately detect drug-resistant strains of the hepatitis C virus.
Researchers Seek Water Test for Invasive Species Detection
Detecting invasive lake and river species using just a water sample would be a dream come true for wildlife managers and regulators in the state and University of Maine researchers may soon make this an inexpensive reality.
New Cell Structure Finding Might Lead to Novel Cancer Therapies
University of Warwick scientists in the U.K. say they have discovered a cell structure which could help researchers understand why some cancers develop.
Ebola Assays Compared in Head-to-Head Analysis
A newly published study has attempted to rigorously evaluate a few of the assays recently granted Emergency Use Authorization by the US Food and Drug Administration to test for Ebola Zaire virus.
Profiling DNA Viruses in Arctic Lakes
The Arctic's freshwater lakes contain viral communities composed of DNA viruses from lineages that are largely distinct from those described elsewhere, a new study suggests.
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