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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.

The synthesis of genetic constructs via the Slonomics™ technology utilizes various liquid handling, mixing, incubation and microplate handling procedures, performed via a multistep, convergent synthesis protocol. For the automated parallel production of such custom gene constructs, we developed a modular system architecture consisting of independently operating robotic stations.

Synthetic biology allows for the direct chemical synthesis of any gene, operon and chromosome. Not only does synthetic biology facilitate copying and editing existing genetic information, it also instantly permits the creation of any imaginable completely novel genetic information without being constrained by nature.

In order to identify host genes required for M. tuberculosis infection and persistence, we developed a phenotypic cell-based assay in both murine and human cells and adapted it for high throughput and high content screening purposes. Knock-down efficiencies above 80% were achieved in “hard-to-Transfect” macrophage cells. Validation of the assay performed with control siRNAs will be discussed.

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

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

The purpose of our study was to evaluate soluble matrixmetalloproteinases (MMPs) and their inhibitor (TIMP1) associated to leg ulcer wound fluid in comparison with the tissue localized ones. We aimed to develop an easy-to-perform protocol for the quantification of soluble MMPs / TIMP and associate the obtained levels with the disease stage and prognostic.

Plasma proteins serve as good indicators of disease as there are representative proteins from several cellular processes and thus a potential source for biomarker discovery. The large dynamic range of plasma proteins makes the analysis very challenging, as a large number of low abundance proteins are masked by a few high abundance proteins.

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.

Protein was extracted from 7-year old FFPE samples of B-cell lymphoma and mamma carcinoma, followed by ULS labeling. Samples were used for single or two color assays using a custom-made antibody array. We observed differential expression levels for several captured analytes. In B-cell lymphoma, expected high levels of IgM were detected, while high levels of CA19-9, a putative marker, were found for the breast cancer tissue.