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| Application Story: Dr Kiki Tahtis
Dr Kiki Tahtis carries out research in the Molecular Angiogenesis Laboratory, at the Weatherall Institute of Molecular Medicine, University of Oxford, UK. Here, she describes her research and how the introduction of ProbeLibrary™ has benefited this work.
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1. What is your research area?
My current area of research is angiogenesis. The focus of our lab is the identification, by novel bioinformatics analysis, of new endothelial specific genes. I’m involved in the subsequent characterisation of those genes. More specifically I’m trying to determine the expression pattern of a gene, which we have shown to be involved in angiogenesis. I’m also very interested in determining how this gene is regulated.
2. How many people work with Real-Time PCR in your group?
There are 11 people in the group and 6 of us work with the ProbeLibrary™. However, several people from other labs are also very interested in the ProbeLibrary™. We all use a Corbett thermal cycler with master mix from ABgene. We previously used SYBR Green and pre-validated probes, but now everyone is basically using the ProbeLibrary™ system. It has fitted in smoothly with the previously established Real-Time PCR routines.
3. Why did you choose to test the ProbeLibrary™?
We were looking for a Real-Time PCR system that was quick, specific, gave highly reproducible results and that could be used by the entire lab in a cost-effective way, and this has been achieved. Also, because we work on very novel genes, we found that with some pre-validated probes the company would not reveal the sequence used in probe design, which meant that we needed to check that we did indeed have a probe to our gene of interest (by cloning). This problem was eliminated by switching to ProbeLibrary™.
4. What have been the main benefits of the ProbeLibrary™?
It has sped up our research and it also saves time in assay design. The system is very convenient because we already have all of the probes we need available in the lab, so we can quickly test several probe/primer sets for a gene of interest and select the best one. It also means that we can easily and cost-effectively create a large bank of house-keeping genes that can be accessed by the lab. We now run almost 50% more assays than previously and we are also considering purchasing a new thermal cycler.
5. Can you disclose your near future research plans?
We plan to use the Human ProbeLibrary™ to characterise the expression levels of recently identified novel endothelial markers using human tumour and normal clinical samples which are available in the lab. We also now have a large bank of cDNA from endothelial cells which have been exposed to a variety of conditions known to regulate angiogenic markers, this will hopefully give us insight as to how some of the genes we are working on are regulated.
With the Mouse ProbeLibrary™, we would like to determine the tumour xenograft as well as the normal tissue distribution of novel endothelial markers in mice, for targeting purposes.
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