Posts tagged ‘PrimePCR’

Bio-Rad Introduces Wet-Lab Validated Real-Time PCR Assays for Rat Genome

Bio-Rad Laboratories announced the launch of new PrimePCR™ Rat Genome Assays for quantitative real-time PCR (qPCR) (available in U.S., Canada, Australia, New Zealand, Singapore, and E.U. only). This expands Bio-Rad’s line of wet-lab validated assays that includes human and mouse genome assays.

Assay validation is one of the most important steps in qPCR but is sometimes ignored because researchers may be unaware of its critical importance in generating accurate results. Validating assays can be laborious and time-consuming, especially when they do not work and need to be redesigned and reordered. While some companies claim to sell prevalidated assays, this assertion often refers to software predictions as opposed to wet-lab validation.

Bio-Rad’s PrimePCR Assays, however, are fully wet-lab validated for specificity, efficiency, and sensitivity, and help researchers adhere to industry best practices known as MIQE (minimum information for publication of quantitative real-time PCR experiments). As part of this validation process, Bio-Rad scientists validate all PCR products using next-generation sequencing, verifying the percentage of on-target amplification. In addition, all of the validation raw data are available to the customer.

Meredith Tennis, PhD, a researcher at the University of Colorado Cancer Center, has been using PrimePCR Assays for cancer chemotherapy prevention studies.

“We choose PrimePCR Assays because they are expertly validated, helping us meet MIQE guidelines,” said Tennis. “Through PrimePCR, we can confidently assess every gene we are interested in on the first try, saving us both time and money.”

Researchers will be able to order PrimePCR Rat Genome Assays as individual assays (desalted or HPLC-purified), or build a custom plate (96- or 384-well). Assays are available for more than 20,000 rat genes as SYBR®- and probe-based assays with a choice of five different fluorophores.

In addition, SYBR® and probe PrimePCR PreAmp Assays will be available for the rat genome. These preamplification assays allow researchers who are working with limited samples to look at more targets during their qPCR experiments. Bio-Rad’s PreAmp Supermix, along with PrimePCR PreAmp Assays, produces consistent gene expression results while delivering the industry’s lowest bias.

For more information on Bio-Rad’s PrimePCR products, please visit http://www.bio-rad.com/PrimePCRpr.

October 15, 2014 at 8:38 am Leave a comment

Bio-Rad Expands Portfolio of PrimePCR™ Assays for Droplet Digital™ PCR

PrimePCR ddPCR imageBio-Rad Laboratories, Inc. announces an expansion of its line of PrimePCR probe assays for Droplet Digital PCR that adds another 56 assays for mutation detection and 109 assays for copy number variation (CNV) analysis to Bio-Rad’s catalog of existing assays for use on its QX100™ and QX200™ Droplet Digital PCR systems.

The new Droplet Digital PCR (ddPCR™) assays are the only available assays that are predesigned and fully wet-lab validated for digital PCR, which frees researchers from time-consuming and labor-intensive assay design and experiment optimization.

Many conventional methods, such as qPCR and next-generation sequencing, offer poor selectivity, fail to detect events with frequencies of less than one in 100 wild-type sequences, and lack the resolution needed to accurately analyze CNV. However, Bio-Rad’s ddPCR technology provides an absolute measure of target DNA molecules and, together with PrimePCR mutation detection assays, can enable detection of one mutant molecule in a background of 2,000 wild-type molecules in a single well without the need for a standard curve. Measuring these extremely low levels of mutation abundance could lead to the development of new diagnostics that are less invasive and dramatically more sensitive. In addition, the high precision and absolute quantification made possible by PrimePCR ddPCR assays enable the quantitative discrimination required to resolve small fold changes in gene copy numbers for CNV analysis.

Professor Per Guldberg, a group leader at the Danish Cancer Society, has been using PrimePCR ddPCR assays to detect mutations in neoplastic cells from body fluids. The assays are helping his lab uncover oncogenic biomarkers for bladder cancer, malignant melanoma, and other types of cancers.

“We have found the PrimePCR digital PCR assays to be very robust and specific with a high signal-to-noise ratio,” said Guldberg. “They have also helped us by entirely eliminating the need for cumbersome assay design and optimization.”

The mutation detection assays include those for deletions in the anaplastic lymphoma kinase (ALK) gene, which are believed to be associated with acquired resistance to ALK tyrosine kinase inhibitor (TKI) cancer therapy. The assays also target mutations associated with other cancers such as pancreatic cancer, melanoma, and lymphomas. Many of the CNV assay targets are associated with cancers (leukemia, melanoma, and uveal melanoma) and neurological diseases.

PrimePCR digital PCR assays are available in multiple reaction sizes and are compatible with both the QX100 and QX200 ddPCR systems using the ddPCR supermix for probes (no dUTP).

For more information on Bio-Rad’s PrimePCR products, please visit www.bio-rad.com/PrimePCR.

May 22, 2014 at 3:14 pm Leave a comment

Bio-Rad Offers New PrimePCR™ Disease Panels for Real-Time PCR

Bio-Rad Laboratories, Inc. announced the launch of more than 700 new PrimePCR human and mouse disease panels and 71 new pathway panels for real-time PCR (qPCR). The panels allow researchers to thoroughly and efficiently investigate genes known to be differentially expressed in a specified pathology or biological pathway. Bio-Rad now offers more than 1,100 panels with nearly 6,700 predesigned plate configurations, establishing Bio-Rad as the industry’s largest and most up-to-date provider of predesigned plates.

The expanded product line includes highly studied pathways such as Hedgehog and bone morphogenetic protein (BMP) signaling, as well as processes such as circadian rhythms and visual perception. In addition, disease categories span hundreds of conditions, ranging from specific cancers and neoplasms to metabolic diseases such as obesity and diabetes.

Working in collaboration with Thomson Reuters, Bio-Rad developed a plate design strategy that ensures that the most relevant gene targets are chosen for each real-time PCR panel; gene targets are weighted for differentially expressed genes and the frequency with which they appear in the peer-reviewed literature. For disease panels, Thomson Reuters referenced the diseases and their corresponding gene targets in the U.S. National Library of Medicine database.

All of the assays were designed strictly adhering to MIQE guidelines: maximum transcript coverage, minimal overlap with known single nucleotide polymorphisms, and spanning large introns where possible. In addition, all of the primer assays have been fully wet-lab validated for specificity, efficiency, and sensitivity to an unprecedented level of quality, according to Jan Hellemans, cofounder of the qPCR and bioinformatics firm Biogazelle, which worked with Bio-Rad to develop these assays.

Bio-Rad has also updated and expanded its existing predesigned pathway and collection panels to include the most scientifically relevant gene targets based on current literature and the latest bioinformatics data.

For more information on Bio-Rad’s PrimePCR products, please visit www.bio-rad.com/PrimePCR.

January 20, 2014 at 3:25 pm Leave a comment

Bio-Rad Introduces New Digital PCR Assays for Human Copy Number Variation and Mutation Detection

PrimePCR ddPCR imageBio-Rad Laboratories, Inc. today announced the launch of its PrimePCR™ assays for Droplet Digital™ PCR. These new predesigned assays for mutation detection and copy number variation (CNV) have been experimentally validated to provide single-copy PCR resolution without a standard curve.

Droplet Digital PCR (ddPCR™) technology provides an absolute measure of target DNA molecules. Researchers can use ddPCR assays to accelerate discovery and enable new research strategies for inherited disorders, cancer, and infectious diseases.

“We are excited because these are the first assays that have been made available as fully validated for digital PCR,” said Richard Kurtz, marketing manager of the Gene Expression Division at Bio-Rad. “These predesigned assays remove the burden of assay design and make digital PCR even easier for our customers.”

In this first release, Bio-Rad offers its customers 14 mutation detection assays with matching wild-type assays for biologically relevant and highly prevalent cancer mutation targets chosen from the COSMIC database. Commonly studied cancer gene mutations such as BRAF V600E and EGFR T790M are among the first set of assays available.

The assays, combined with the sensitivity of Bio-Rad’s ddPCR system, are capable of detecting a single mutant copy in a background of 2,000 or more wild-type molecules (0.05% mutation frequency). The precision of ddPCR also enables cancer researchers to discriminate small-fold copy number changes with the 62 assays targeting common cancer genes and two reference target assays now available.

Bio-Rad has partnered with Biogazelle, a qPCR data analysis and services company, and Integrated DNA Technologies (IDT), a world-class manufacturer of oligonucleotides, to design, optimize, and experimentally validate the assays. The PrimePCR ddPCR assays employ universal cycling conditions and do not require optimization. Primer specificity has been validated by next-generation sequencing, and all assays have been validated on Bio-Rad’s QX100™ Droplet Digital PCR system.

PrimePCR ddPCR assays are available in multiple reaction sizes.

For more information on Bio-Rad’s PrimePCR products, please visit www.bio-rad.com/PrimePCR.

July 11, 2013 at 3:09 pm Leave a comment

PrimePCR™ Pathway Analysis: Pathway Curation and Real-Time PCR Panel Design Strategy

Pathway analysis is an efficient approach for studying a large number of related gene targets in a single experiment. Bio-Rad Laboratories and Thomson Reuters, a leading provider of system biology tools, have partnered to provide predesigned pathway panels for gene expression analyses using real-time qPCR. This tech report describes how Bio-Rad’s PrimePCR™ pathway panels were curated and designed using a ranking strategy developed by Thomson Reuters.

Read more…

April 8, 2013 at 3:21 pm Leave a comment

PrimePCR and MIQE

The MIQE guidelines have set a new standard for publishing qPCR results to ensure integrity in the scientific literature and increase experimental transparency. One of the key guidelines focuses on the proper design and experimental validation of primer assays used in qPCR. This tech report describes the design and wet-lab validation of PrimePCR assays and how they comply with the MIQE guidelines.

Read PrimePCR™ Assays: Meeting the MIQE Guidelines by Full Wet-Lab Validation.

January 23, 2013 at 4:52 pm Leave a comment

Building a custom PCR assay that conforms to MIQE guidelines

If you’ve been following the news lately, you’ve likely heard about Bio-Rad’s new PrimePCR platform which provides scientists with commercial qPCR kits that are guaranteed to conform to the MIQE guidelines. If you haven’t heard about the PrimePCR platform then you can learn more by reading the article MIQE trouble free posted on the Canadian Biotechnologist.

Below we present you with a video tutorial on how you can use the PrimePCR platform to build a custom assay.

November 19, 2012 at 3:29 pm Leave a comment

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