Posts Tagged ‘qPCR’

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RT-PCR Directly from Cultured Cells

 :: Posted by American Biotechnologist on 09-08-2014

Bio-Rad Laboratories announced the launch of new rapid cell lysis kits that allow researchers to obtain reverse transcription quantitative PCR (RT-qPCR) data directly from cultured cells without the need for a separate RNA purification step. Bio-Rad’s SingleShot™ family of cell lysis RT-qPCR kits provide high-quality gene expression results in less than two hours.

Available column isolation methods for purifying RNA are time-consuming and laborious while other methods that enable RT-qPCR directly from cell lysates can damage the RNA and result in poor genomic DNA clearance. However, Bio-Rad’s SingleShot Kits eliminate those challenges and offer superior reproducibility and accuracy of gene expression results. In addition, minimal setup and pipetting steps create an automation-friendly workflow and, unlike other similar methods, Bio-Rad’s kits do not require an additional pipetting step to stop the cell lysis reaction.

 

The SingleShot Kit removed genomic DNA (Cq > 40) while another available kit resulted in poor genomic DNA clearance.

The SingleShot Kit removed genomic DNA (Cq > 40) while another available kit resulted in poor genomic DNA clearance.

 

“The SingleShot Kits are ideally suited for high-throughput laboratories with large-volume workloads and for researchers who are faced with a limited number of cells and require extreme accuracy in each analysis,” said Paul Streng, senior product manager in the Gene Expression Division of Bio-Rad’s Life Science Group.

Bio-Rad’s SingleShot Kits are the only available kits that include an RNA control template and qPCR assay to help researchers determine optimal cell number and lysate inputs for their RT-qPCR reactions.

SingleShot Kits are validated for use with a wide variety of adherent and suspension cell lines and are offered in multiple formats, including one-step RT-qPCR and two-step RT-qPCR kits that are compatible with either SYBR® Green or probe-based assays. SingleShot Kits are available as a stand-alone cell lysis kit. All SingleShot Kits are validated for use with PrimePCR™ Assays and Panels.

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Overcoming PCR Inhibitors

 :: Posted by American Biotechnologist on 07-15-2014

Sso advanced universal inhibitorBio-Rad Laboratories announced the launch of the SsoAdvanced™ Universal Inhibitor-Tolerant SYBR® Green Supermix, a proven solution for obtaining reproducible, optimal-quality qPCR data from the most challenging samples.

Generating quality qPCR data from difficult-to-amplify samples such as crude lysates from plants, tissues, formalin-fixed paraffin-embedded samples, and other less-than-ideal sources that contain PCR inhibitors can be challenging and frustrating. In these situations, PCR may be greatly inhibited, making it nearly impossible for researchers to obtain reliable data, wasting time and money.

The formulation of the SsoAdvanced™ Universal Inhibitor-Tolerant SYBR® Green Supermix has been optimized and validated to tolerate a wider spectrum of PCR inhibitors than competing SYBR® Green reagents. These inhibitors include those found in crude lysates, polysaccharides and polyphenols, and various reagents left over from sample prep such as ethanol, isopropanol, EDTA, and sodium chloride see performance data for all inhibitors.

As with all SsoAdvanced Supermixes, the inhibitor-tolerant version includes Bio-Rad’s Sso7d-fusion polymerase, which is engineered for enhanced qPCR performance. The supermix is compatible with all real-time PCR instruments and functions under any reaction condition.

Visit www.bio-rad.com/SUIT1 to learn more about SsoAdvanced™ Universal Inhibitor-Tolerant SYBR® Green Supermix.

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Improving Visibility of qPCR Reaction Mixtures

 :: Posted by American Biotechnologist on 05-14-2014

Bio-Rad Laboratories recently announced the launch of its Precision Blue real-time PCR dye, making it easier for researchers to set up real-time PCR (qPCR) experiments.

The blue dye improves the visibility of qPCR reaction mixtures, enabling loading of small volume reactions with greater speed, accuracy, and precision. Applications such as single nucleotide polymorphism (SNP) genotyping and high-throughput gene expression analysis using 384-well plates will particularly benefit from use of this dye.

Precision Blue dye is compatible with Bio-Rad’s universal qPCR supermixes (iTaq™ universal and SsoAdvanced™ universal supermixes) and does not interfere with SYBR® Green- or probe-based detection. The dye can be used with all Bio-Rad CFX real-time PCR systems and all other vendors’ real-time PCR systems.

For convenience, Precision Blue can be added directly to the supermix or used as a component of a reaction master mix. The dye is concentrated 200x, and delivers 500 x 20 µl reactions.

For more information about Precision Blue dye, please visit www.bio-rad.com/PrecisionBlue.

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Bio-Rad Sponsors New Version of MIQE qPCR App for Real-Time PCR and Digital PCR

 :: Posted by American Biotechnologist on 08-28-2013

miqe appBio-Rad Laboratories is sponsoring a new version of the MIQE qPCR app. Researchers can use the new version to validate their digital PCR (dPCR) experiments according to the recently published digital MIQE (dMIQE) guidelines (Huggett et al. 2013).

Professors Michael Pfaffl (TU Munich, Weihenstephan, Germany) and Afif Nour (King Abdulaziz University, Saudi Arabia) designed the original MIQE qPCR app to help researchers improve their real-time PCR (qPCR) assay protocols by making it easier to adopt a set of best practices described in an earlier publication: The MIQE guidelines: Minimum information for publication of quantitative real-time PCR experiments (Bustin et al. 2009). Progress bars in the app show the percentage of assay compliance as each item in the MIQE checklist is completed.

“We were pleased to find that the MIQE qPCR app encouraged our customers to follow the new MIQE guidelines,” said Jean-Pierre Dakkak, a laboratory equipment trading company manager.

“Researchers are really eager to learn how this app can make their life easier,” said Dr. Nour. “They report it instills confidence in the validity of every qPCR or dPCR project.”

The new updates include:

  • Project-specific checklists — checklists remove unnecessary items; for example, reverse transcription items are irrelevant in DNA research
  • Updated references — researchers can stay current with the latest MIQE and dMIQE literature and qPCR symposiums
  • Easy export — users can save their projects and share them with colleagues

The MIQE qPCR app runs on the Apple iPhone, iPod touch, and iPad. It has been downloaded more than 6,500 times in 87 countries. To get your copy, visit http://bit.ly/MIQEapp.

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Innovative qPCR Reagents for Superior Performance

 :: Posted by American Biotechnologist on 04-29-2013

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