Real-time qPCR/PCR

Incorporating preamplification into your qPCR experiments may seem daunting, but it is a powerful way to analyze targets from limited samples. The tips in this article will help you understand how preamplification can enhance your qPCR experiments and the considerations and implications for incorporating it into your workflow.

Comprehensive and effective gene expression studies evaluate both transcriptional and translational regulation. A streamlined protocol was developed for the parallel analysis of mRNA and protein levels using a single lysate produced from cultured cells.

It is imperative to analyze both the transcriptional and translational profiles for a complete understanding of the functioning of genes. Here we describe a simple protocol that enables parallel analysis of both RNA and protein from single cell culture lysates. See how this workflow could save time and money and yield more reliable data than the traditional protocols used.

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.

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.

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.

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.

Traditional real-time PCR workflows often limit the number of genes that can be studied, especially when sample amounts are small. A new workflow now allows Mark Kibschull of the Lunenfeld-Tanenbaum Research Institute in Toronto to track the expression of hundreds of genes using a fraction of the starting material required by traditional methods.

Conducting a triplex one-step RT-qPCR assay promises a more efficient and economical analysis of gene expression compared to running three individual assays. This technical report describes a colorectal cancer case study where a triplex multiplex assay was validated.