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Improving the efficiency of 384 "mini-tube" technology using mosquito nanlolitre pipetting
Joby Jenkins, Wayne Bowen, Rob Lewis & Chloe Milburn

384-well plate “mini-tube” consumables have been developed by automation companies such as REMP and The Automation Partnership (TAP) to hold compound samples divided into single-use storage tubes. Each tube holds 40 -75 µL, and is filled, sealed, stored and then thawed just once before use. However, when accessed by conventional pipetting technology, the tube’s practical working volume is as little as 20 µL, which means high compound wastage. The seal must also be pierced prior to pipetting to p

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Application of genetic programming in analysis of quantitative gene expression profiles for identification of nodal status in bladder cancer
Anirban P. Mitra, Arpit A. Almal, Ben George, David W. Fry, Peter F. Lenehan, Vincenzo Pagliarulo, Richard J. Cote, Ram H. Datar, William P. Worzel

Nodal involvement in bladder cancer is an independent indicator of prognosis. This study employed an iterative machine learning process called genetic programming on quantitative expression values of 70 genes to classify primary urothelial carcinoma samples into those associated with or without nodal metastasis. The generated rules showed a strong predilection for ICAM1, MAP2K6 and KDR resulting in gene expression motifs that cumulatively suggested a pattern ICAM1>MAP2K6>KDR for node positive ca

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How to Perform High Throughput siRNA Transfection
Mark Hewitson, Hanno Hermann and Neil Benn

We demonstrate the optimization of siRNA transfection process for time and costs in a high throughput application with two independent measurements of the RNAi effect: The visual monitoring of Eg5 expression and the quantitative PCR measurement of GAPDH mRNA after transfection of Hela cells. The results show, that 1 wash cycle is sufficient for the removal of any remnants of the transfection complex from disposable tips, making the tips re-usable.

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Quantification of siRNA by a Novel Competitive-qPCR Method
Wei-li, Liu., Mark Stevenson and Len Seymour

We have developed a competitive qPCR method in which siRNA competes with a homologous forward primer to bind template DNA, giving siRNA concentration dependent inhibition. The addition of E6-siRNA to cqPCR led to inhibition of amplification in a linear concentration-dependent manner, with as little as 200pg of siRNA capable of being detected. Irrelevant siRNA had no effect on amplification confirming specificity

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Quantification of siRNA by a Novel Competitive-qPCR Method
Wei-li, Liu., Mark Stevenson and Len Seymour

We have developed a competitive qPCR method in which siRNA competes with a homologous forward primer to bind template DNA, giving siRNA concentration dependent inhibition. The addition of E6-siRNA to cqPCR led to inhibition of amplification in a linear concentration-dependent manner, with as little as 200pg of siRNA capable of being detected. Irrelevant siRNA had no effect on amplification confirming specificity

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The Silencing of Multidrug Resistance-Associated Protein 5 by siRNA Complexes
M.Malinen, E.Mannermaa, T.Ryhänen, E.Kuusela, M.Häkli, M.Yliperttula and A.Urtti

The purpose is to study the role of multidrug resistance-associated protein 5, MRP5 in drug metabolism in human retinal pigment epithelium (RPE). RPE forms the outer part of blood-retinal barrier (BRB) which restricts movements of solutes from systemic bloodstream to the neural retina. The efflux protein, MPR5 is expressed in RPE but its functions are mainly unknown. SiRNA will be tested as a tool to clarify the role of MRP5.

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Caveolin-1 Expression as a Possible Biomarker in Pancreatic Cancer Diagnosis
C. Tanase, E. Raducan, L. Albulescu, E. Codorean, M.I. Nicolescu, D.I. Popescu, M.L. Cruceru and A.C. Popa

Caveolin1 (Cav-1) function either as a tumor supressor or as a promoter of metastasis. Overexpresion of cav-1 was correlated with: tumoral grading, proliferration markers (Ki67, p53), serum tumor markers (CEA, CA19.9) and angiogenic markers (VEGF, bFGF).

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New Transcripts Identified for S. cerevisiae, S. pombe and Drosophila Using Novel cDNA Cloning
Les Hoffman, Janina Gornemann, Erica Rodriguez and David Brow

Unannotated transcripts, including antisense, noncoding, intergenic, and potential regulatory RNAs, were identified in three different eukaryotic model organisms by cDNA cloning and sequencing. A rapid double-stranded cDNA generation system will be described.

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Preliminary Report: The Geriatric Propamed Study: Prospective pharmacopgenetics in geriatrics
Dr LS Griffith, Dr L Chialda and Dr A Pahl

In a worldwide first proscpective phamacogenetic study preliminary results show reduction of adverse drug reactions and hospitilisation stays for geriatric patients after pharmacogenetic testing and medication interaction analysis to fit the medicine to the patient.

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Scientific News
CDC Develops a New, Faster Lab Test for Enterovirus D68
Confirmed cases will appear to rise as agency accelerates specimen testing.
NanoString, Brigham and Women’s Hospital Collaborate
Research collaboration to accelerate translation of genomic discoveries into clinical diagnostics in oncology.
Unlocking The Non-Coding Half of Human Genome
Texas A&M biologists unlock non-coding half of human genome with novel DNA sequencing technique.
Improving Trout Resistance to a Deadly Disease
A highly sensitive real-time PCR test has been developed to measure F. psychrophilum in fish tissue.
Blood Test To Diagnose Depression
Test identifies nine blood markers tied to depression; predicts who will benefit from therapy.
Redefining High Risk Patients With Stage II Colon Cancer
microRNA-21 identified as an independent prognostic biomarker.
More People Testing Positive for Chikungunya Virus in the U.S.
Clinical testing volume and positivity rate both grew after the first case of the infectious disease spread to Western Hemisphere this year.
Novel rhPCR Unravels Gene Expression
Method enables identification and analysis of highly similar alternatively spliced transcripts.
Detecting and Identifying Candida Species in Blood Samples of Critically Ill Paediatric Patients
The study aimed to develop a multiplex nested PCR method to detect and identify seven Candida species in peripheral blood samples of critically ill paediatric patients.
IDT Shares Comprehensive qPCR Resources
Special qPCR compendium issue of DECODED now freely available.
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