|NEW TECHNOLOGIES FOR FFPE SAMPLES: Improved RNA Isolation and novel cDNA priming for qPCR and for universal mRNA amplification|
G Krupp1; R Jaggi2; D Englert3, DJ Wilson3, S Laken3, S, ES Quabius4
Improved FFPE RNA isolation, novel TR priming for complete recovery of FFPE mRNA fragments. Differential expression defines Scores for tumor classification: comparing fresh-frozen with FFPE. Novel flow-through Ziplex microarray platform demonstrates MAQC performance level of data obtained with FFPE samples.
|FOXP3 Gene Expression in Multiple Sclerosis patients before and after Mesenchymal Stem Cell therapy|
Maryam Mohajeri, Mandana Mohyeddin Bonab, Behrooz Nikbin, Ali Farazmand
Multiple Sclerosis (MS) is a chronic inflammatory demyelinating disorder of the CNS. No successful treatment for MS, but one therapeutic strategy in research is the use of bone marrow-derived mesenchymal stem cells (MSC). We studied a group of MS patients who underwented MSCs, assayed for expression of a transcription factor, FOXP3, as a specific marker of MS amelioration in peripheral blood. qRT-PCR on PBMCs showed higher FoxP3 levels.
|miR-21 upregulation and miR-128a downregulation in human glioblastomas|
P. Costa1,2, A. Cardoso1, L. Almeida1,3, M.C. Pedroso de Lima1,2, P. Canoll4, J. Bruce5
miRNA deregulation is an important hallmark on cancer development and progression. This work shows that miR-21 is overexpressed is human glioblastoma samples and cell lines, whereas miR-128a is highly downregulated on these tumours. miRNA modulation using antisense oligonucleotides and/or miRNA precursor molecules decreased tumor cell viability. Thus, targeting deregulated miRNAs may constitute a complementary/alternative approach on the treatment of glioblastomas and other miRNA-related maligna
|Partial characterization of nitrate signaling pathway of Arthrospira platensis PCC 7345 |
Pradeep kumar L, and Vani B.
The present study is to elucidate the signaling network involved in nitrate uptake and assimilation of Arthrospira platensis. Expression of adenylate cyclase, cyaC was found to be nitrate signal dependent. The involvement cAMP was also observed in different levels of nitrate signaled assimilatory pathway. We have used differential display to identify a histidine kinases in the nitrate signaling of Arthrospira platensis.
|Chemically Modified Primers for Improved Multiplexed 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. Furthermore, preferential amplification of certain targets leads to an unequal distribution of amplicon products, making quantifi
|Hot Start dNTPs - A Novel Tool for Controlled Nucleotide Incorporation in PCR|
Tony Le, Elena Hidalgo Ashrafi, Sabrina Shore, Victor Timoshchuk, Natasha Paul, Richard Hogrefe, Inna Koukhareva, Alexandre Lebedev
PCR is a widely used scientific tool employed by a variety of applications. Various Hot Start technologies have already been developed using modified PCR components to increase specificity of a reaction. Recently developed CleanAmpTM dNTPs are modified nucleoside triphosphates with a thermolabile 3’-tetrahydrofuranyl protecting group that is released at higher temperatures. These modified dNTPs prevent low temperature primer extension, which can often be a significant problem in PCR. At higher t
|Evaluation of microfluidic digital PCR for the detection of cancer biomarkers|
Rebecca Sanders, Claire Bushell, Carole Foy, Daniel J. Scott
dPCR is achieved by sample partitioning prior to PCR amplification such that each reaction chamber contains one copy or less of target DNA. This dilution becomes the limiting factor and an accurate target molecule count is achievable. This study evaluates dPCR’s quantitative capabilities and investigates parameters influencing copy number quantification, using the Fluidigm Biomark instrument. Biomark technology combines dPCR theory with a microfluidics platform.
|Inhibition of DNA methylation does not overcome docetaxel resistance in human breast cancer cells|
Kastl L, Brown I, Schofield A
DNA mehtylation can lead to chemotherapy resistance in cancer. The aim of this study was to investigate the role of DNA methylation in docetaxel-resistant human breast cancer cells. Whereas the DNA methylation machinery is altered in docetaxel-resistant human breast cancer cells compared to docetaxel-sensitive human breast cancer cells, resistance to docetaxel could not be reversed using the DNA methylation inhibitor decitabine.
|A novel multiplex qPCR for the simultaneous detection of Salmonella spp., E. coli O157:H7 and Listeria monocytogenes from raw Atlantic Salmon fillets|
Sonja S. Nitecki, Brian F. Carney, John W. Slater, Wolfram M. Bruck
A triplex qPCR assay was developed to screen for three major food-borne pathogens in fish. The assay is based on simultaneous qPCR amplification of specific target genomes using fluorophore labelled locked nucleic acid (LNA) TaqMan probes. The detection limit of the qPCR is 1 colony forming unit (cfu) per 1ml and the detection limit of the assay with 24h general enrichment is 1 cfu per 25g of fish meat.