Molecular Core Online Order

The goal of this service is to give the basic science research community access to quantitative PCR in a timely and cost effective manner.

The core has CFX96 from BioRad and associated software and has accumulated sufficient experience to give direction to the uninitiated investigator in their use. Along with a Zymark automated loader once you have tested your experimental system, set-up 4 96-plates drop off the plates at the Core and get your results next day.

We strongly recommend that a new users plan their first experiments with the assistance of the core staff or a user who has been trained on the CFX96 so one can learn how to use the software and appropriate reaction conditions. You will need access to a PC computer to operate the software that sets the PCR running conditions for the machine and creates a plate set-up so that data is collected from the correct wells. The plate set-up will also be used in the downstream analysis of the data. Mac users can purchase PC emulation software that will run this program satisfactorily.

In practice, an investigator will need to decide whether they will do a Relative Quantitification (RQ) study or Absolute Quantification (AQ) and then if they will use primer probe system (Taqman assays) or SYBR Green as a detection. Currently Applied Biosystems, SABioscience through Qiagen, IDT, BioRad other vendors make available pre-designed primers with established running conditions for many genes that will save time. If you decide to design primers or use primers that have been established for regular PCR, take into consideration that optimization for preventing primer dimers, similar annealing temperature, cDNA/mRNA specificity and efficiency of 95% or greater are important for a successful qPCR experiment.

Prior to bringing the plate to the Molecular Core, you need to register a request using Online Ordering System and send email with the running conditions to Molecular Core. Set up your 96 well plate and then drop it off at the Molecular Core. The core staff is available to answer most questions and the core stocks the kits and enzymes that are commonly used for qPCR. In order to cover the service contracts of these machines we charge a $15 per run fee, reguardless of the failure or success of the run.