Taqman Core Facility

Animation of a Taqman PCR reaction. Polymerisation from the primers forces the cleavage of the probe. The Reporter dye is freed from its quencher and fluoresces when excited by a laser.
Animation of a Taqman PCR reaction. Polymerisation from the primers forces the cleavage of the probe. The Reporter dye is freed from its quencher and fluoresces when excited by a laser.

One of the key challenges in developing gene transfer technologies is to be able to detect and quantify the levels of gene expression from our vectrs in-vivo. In order to do this we rely on Taqman RT-PCR to quantify levels of vector derived mRNA. Here in Oxford we run a core Taqman facility for the whole Consortium.

 

 

 

 

 

Our Biomek NX automated liquid handling 'robot' and one of our 7900 Real-time PCR machines.
Our Biomek NX automated liquid handling 'robot' and one of our 7900 Real-time PCR machines.

Experiments are carried out all over the consortium and samples are sent to us for processing. This process is largely automated as we make use of an automated liquid handling workstation to purify samples and setup reactions plates. This leads to far greater accuracy, precision and a higher throughput that could be achieved manually. The vast majority of the assays we used a developed and optimised in-house. Assays that we developed for our own lab work are now routinely used to detect expression in our clinical trial samples from the ongoing clinical programme.

The levels of fluorescence produced by the PCR reaction can be used to quantify the amount of material that was present at the start. Hence the term, quantitative PCR.
The levels of fluorescence produced by the PCR reaction can be used to quantify the amount of material that was present at the start. Hence the term, quantitative PCR.

We are fortunate to have two ABI 7900 Taqman machines to carry out all our Taqman work on. These machines are capable of running many hundreds of samples a day.

For data analysis we have developed a range of proprietary software tools in order to streamline and standardise the process.

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