Designing a Quantitative Western Blot Experiment to Avoid Common Pitfalls

The most important video you will ever watch for performing proper RT-qPCR

Did you think that sample prep and primer design was straight forward? What impact can the use of different housekeeping genes as controls have on your results? You will be shocked at what you find out.

In this video, Sean Taylor of Bio-Rad Laboratories explains exactly what you must know to perform RT-qPCR according to MIQE standards.

How to ensure significant results from your qPCR studies

So you’ve heard all the hoopla about MIQE and how important it is to follow the guidelines when conducting your real time qPCR experiments, (if you haven’t, you better check this out!), but where’s the proof that following MIQE actually makes a difference? After all, qPCR was around for several years before anyone came up with this MIQE stuff. Right? Well…maybe not! As it turns out, qPCR experiments that don’t follow MIQE guidelines can be very difficult for others to reproduce and can even lead to incorrect conclusions in gene expression studies.

In a recently released case study involving breast cancer patients, researchers found that the MIQE guidelines played a central role in obtaining the expected conclusions with a positive control target. The article was written to show readers in a simple, stepwise process how to design a good qPCR experiment that covers the major components of the MIQE guidelines. While each step of the experimental design was found to impact the final conclusion (sample collection, RNA quality and purity and the use of appropriate primers), the most striking result was the impact of reference gene selection on the results. At one extreme, normalization by the commonly used GAPDH and 18S reference genes gave either no significant results of statistically significant data that was opposite to the expected outcome, while other more stable reference genes, (HPRT1 and TBP), gave statistically significant data that supported the conclusions from previously published results with this target.

The study concludes that the application of the MIQE guidelines to qPCR experiments result in reliable, quantifiable and reproducible data. With a growing list of journals that are now requiring the submission of supplemental data supporting adherence to the MIQE guidelines,the publication of qPCR data will become more challenging if they are ignored.

So don’t miss out on significant data. Use MIQE!

To read the case study click here.

An Advanced Look at Droplet Digital PCR

As a follow up to our video introduction to droplet digital PCR, we are proud to present you with an advanced video presentation on droplet digital PCR technology. Please have a look and let us know what you think!

Video Introduction to Droplet Digital PCR

As promised in our earlier post on Droplet Digital PCR, we are proud to present the following educational training video “An Introduction to Droplet Digital PCR”, given by award winning field application specialist Dr. Sean Taylor.

Be sure to stay tuned for the advanced training video which will be posted in the near future.