It’s no surprise that Droplet Digital PCR (ddPCR) has found its place in the world of cancer research. Bio-Rad’s ddPCR™ technology has a remarkable ability to quantify miniscule amounts of target DNA and RNA, and thus can contribute to early detection of rare tumorigenic mutations against a high background of “normal” DNA as well as to other applications, including identifying cancer subtypes, optimizing drug treatment plans, and studying tumor evolution. At the Fred Hutchinson Cancer Research Center in Seattle, scientists are producing some of the most important advances in prevention, early detection and treatment of cancer and other diseases. They are studying the disease process from every angle seeking to uncover factors that influence a person’s likelihood of getting cancer. Understanding of such factors, of course, can help reduce risk and save lives. To learn more about how Droplet Digital PCR is being wielded to fight against cancer, we caught up with two researchers – Muneesh Tewari and Jason Bielas— who are using Bio-Rad’s QX100™ Droplet Digital™ PCR system in their quest to break through the current limits of nucleic acid detection and quantification.
Posts Tagged ‘Bio-Rad Laboratories’
Transforming Cancer Research: Droplet Digital™ PCR Plays Integral Role at Fred Hutchinson Cancer Research Center:: Posted by American Biotechnologist on 04-15-2014
Image Lab™Software: Learn the Basics from the Experts
Join Bio-Rad Laboratories for a 30 minute live webinar developed and delivered by Ben Wang, PhD, Senior Technical Support Specialist, Bio-Rad Laboratories
The webinar will take place TOMORROW – Wednesday April 9, 2014 at 9:00 AM Pacific.
As you get ready to use your new system, Bio-Rad will provide you with an opportunity to learn the basic features of Image Lab Software. This training will cover the primary tools, as well as how to acquire an image.
The webinar will be recorded and the playback link will be shared with registrants
Click here to register for the webinar.
A methods article published yesterday provides a rigorous and concise workflow with specific instructions on how to produce and analyze quantitative data using western blot experiments. The paper, coauthored by Bio-Rad scientists and published in BioMed Research International, also highlights recently introduced technologies that improve reproducibility. The result is a powerful, step-by-step guide to obtaining quantitative and reproducible densitometric data from western blots regardless of the specific experiment.
Although western blotting is a well-established laboratory technique, it has recently come under fire as a quantitative method because extreme care must be taken when generating and interpreting the resulting data.
The technique is challenging and requires following a rigorous methodology to achieve reproducible and quantitative data. According to a recent survey of more than 750 labs, 41% of researchers say their western blots fail a quarter of the time.
Dr. Aldrin Gomes, an assistant professor at University of California, Davis, agrees that flawed western blots are not unusual. To compare expression of a protein of interest from sample to sample, protein abundance is commonly normalized to a housekeeping gene. “When I see a large, dense band for the protein of interest or the housekeeping protein, I cringe,” says Gomes. That dense band usually means the protein of interest or housekeeping protein was no longer within the assay’s linear dynamic range. No accurate quantitative data can be extracted from such blots.
Sean Taylor discusses the BioMed Research International paper he coauthored.
Another common reason for failure of quantitative western blots is flawed or incomplete protocols, according to Sean Taylor, the paper’s lead author and a Bio-Rad field application scientist (watch a video of him discussing the paper on the left). To address this, Taylor’s review pays special attention to experimental design and sample preparation and discusses proper definition of the linear dynamic range of protein loading, all key factors for generating meaningful quantitative western blot data.
Taylor also introduces more advanced concepts to improve reproducibility, simplify workflow, and reduce the time and cost of western blotting. One such technique is stain-free total protein normalization, which over the past year has proven superior to using housekeeping proteins or total protein staining to correct for loading errors.
With this article, Taylor hopes researchers now have a simple guide to ensure quantitative and reproducible western blot data for all research fields that rely on this technique.
To read the open access research article, visit http://bit.ly/1kCAOcr.
For additional resources please consult Bio-Rad’s guide to Troubleshooting Western Blots.