Real-time qPCR/PCR

Do we ever think seriously about PCR plates, seals, and tubes as contributing to the quality of our gene expression results? Chances are that these PCR plastics are viewed just as labware by most people. In this article we bring out how simple factors, such as PCR plate rigidity, color of the wells, and adhesiveness of plate seals, can affect your gene expression results and show why it is important to use quality PCR plastics.

Tumor metastasis is a complex process. It requires the ability of the cancer cells to invade their surroundings and travel to distant sites and survive. Dr. Traci Lyons studies the mechanism of COX-2 functioning in the metastasis of breast cancer cells. Find out how Dr. Lyons used Bio-Rad’s PrimePCR™ Assays to confirm COX-2 knockdown at the mRNA level.

Traci Lyons describes how PrimePCR Assays ended her laboratory’s struggle to find a qPCR assay that could reliably detect transcription of the cyclooxygenase-2 (Cox-2) gene, which they had identified as potentially prometastatic. Using the PrimePCR Assay for Cox-2, researchers in her lab were finally able to verify cyclooxygenase-2 knockdown, which is a necessary step toward confirming its role in breast cancer metastasis.

Removing genomic DNA contamination from cDNA preparations is a challenge. All currently available methods are time consuming, could lead to sample loss, and are not efficient. Bio-Rad’s iScript™gDNA Clear cDNA Synthesis Kit tackles these issues and makes this step effortless. This article presents the performance data of the kit and describes how simple and efficient it is to use the kit for gDNA removal.

What’s the optimal melting temperature for a hydrolysis probe? How long should your probe be? And why should you avoid 5′ guanines like the plague? This episode answers these questions and gives you five simple tricks for designing the perfect hydrolysis probe.

During a PCR reaction, it is imperative for the DNA polymerase to remain attached to the template while extending the new strand. This association is jeopardized under challenging PCR conditions and when amplifying longer DNA fragments. See how Sso7d, a small protein capable of binding with the polymerase, helps in stabilizing this association and increasing PCR efficiency.

Follow these six simple tips and you’ll increase the reproducibility of your qPCR experiments.

A qPCR workflow involves multiple steps, each of which adds to the time and cost of the entire experiment. The PrimePCR™ Assays and Panels from Bio-Rad eliminate a number of these steps with their predesigned primers and probes and help with the assay design. See what steps they eliminate and how that would help expedite your qPCR experiment.

Are you using ethanol to prep your bench for qPCR? Watch this episode to find out why that might be a bad idea. We’re sharing seven tips for minimizing contamination of qPCR experiments.

How long should a qPCR primer be and what types of sequences should you avoid? In this episode we share eight tips and tricks for primer design for your qPCR experiments.