Quantitative real time PCR on your behalf
You would like to
quantify genes or their expression, but you are lacking in technical or personnel resources?
Oncoscreen is the solution!
Performance of quantitative PCR and RT-PCR on your behalf
Leave your questions about quantitative analysis of nucleic acids for us to answer. Our molecular biologists with Ph.D qualifications develop solutions tailored to your problems.
Our services are executed using an ABI Prism® 7700 Sequence Detector System with the help of the efficient TaqMan® chemistry.
Take advantage of the expertise of Oncoscreen to accelerate your project.
We accept all kinds of biological samples including paraffin preserved samples or already extracted nucleic acids. With the help of optimised extraction methods, we isolate the nucleic acids for you in constantly high quality – a prerequisite for precise quantitative PCR analyses.
As a result we deliver you an analysis report with graphical presentation, explanations of the experimental protocol and the evaluation of your data.
You can find several applicable examples on the page "Molecular Biological Assisstance of your Clinical Studies" and "Clinical Studies directed by Oncoscreen Ltd. (GmbH)"
Our team remains at your disposal for further questions at all times.
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Raw data for the quantification of two genes (red and green) in four dilution levels (1:2:5) of a cDNA. |
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10 ml-aliquots from the blood of a healthy volunteer were combined with 50 – 100,000 cells of the line HepG2. Afterwards cDNA was extracted and the alpha-Fetoprotein gene (AFP) quantified. AFP is expressed by hepG2 and is hardly detectable in the blood of healthy volunteers. |
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17 different cDNA preparations were produced within four months from the same specimen and the Thymidylate Synthase was quantified. |
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A Special Offer for Newcomers
Are you playing with the thought of setting up TaqMan-PCR in your laboratory, but are hesitant due to the risk of high costs?
Then we have a special offer for you:
- We can develop your first professional primer/probe set (see below which primers are suitable)
- We carry out the first experiments for you, in order to enable you better estimate the projected success of your project.
- We advise you during the introduction of the TaqMan technology in your laboratory.
- Our first measurement results give you concrete standards on which you can orientate yourself
You are completely free to decide at which stage of the project you would like to transfer the technology into your laboratory. We are of course, more than willing to accompany your project in our laboratory to its completion.
Our team remains at your disposal for further questions at all times.
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Which primers are suitable?
The quantitative TaqMan-PCR used by Oncoscreen relies upon that, during the annealing phase a third oligonucleotide binds to the target sequence in a sequence specific manner. This third oligonucleotide (the TaqMan probe) is fluorescently labelled. During the elongation phase the TaqMan probe is hydrolysed by the Taq polymerase, whereby a change in its emission spectrum is used for the quantification.
This complex process requires special reaction conditions. Generally speaking, a melting temperature of 60 °C for the primer and one of 70 °C for the TaqMan probe, are thought to be best. Furthermore, special rules need be observed during the design of the TaqMan probe, and among others, the TaqMan probe should not bind to the target sequence more than 5 nucleotides downstream from a primer. For these reasons, the commonly used PCR primers are unsuitable for the application in the TaqMan PCR.
Oncoscreen has far reaching experience in the field of development and scrutiny of primer/probe sets for the TaqMan PCR. We devise the sets, optimise the reaction conditions and check the efficiency of the PCR according to a standard procedure. The customer receives from us not only an optimised primer/probe set, but also valuable tips and information on future quality control, so that the sets can later be ordered and used even independently of Oncoscreen.
Additionally, the opportunity exists of drawing on a ever increasing number of prefabricated primer/probe sets from Applied Biosystems, the manufacturer and marketer of TaqMan accessories. These are sold under the designation "TaqMan PDAR Target Reagents for Gene Expression". Oncoscreen can also if desired, carry out your order using the TaqMan PDAR Target Reagents. A list of current TaqMan PDAR Target Reagents can be found at the web site of Applied Biosystems: http://www.appliedbiosystems.com
In addition to the deployment of TaqMan probes, we are also competent in quantifying using the Sybr®Green method. Here lower costs, but an increased number of artefacts, are typical, which is why we only recommend this method under special circumstances.
Our team remains at your disposal for further questions at all times.
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TaqMan® and ABI Prism® are registered Trade Marks of the company Applied Biosystems, 850 Lincoln Centre Drive, Foster City, CA 94404, USA Sybr® is a registered Trade Mark of the company Molecular Probes, Inc., PO Box 22010, Eugene, OR 97402-9165, 4849 Pitchford Ave