Posts Tagged ‘Sean Taylor’
The Bio-Plex system is a powerful system for quantitative analysis of up to 100 different proteins, peptides, DNA fragments and RNA fragments from a single drop of sample.
In this video given by Dr. Sean Taylor, you will be given a comprehensive introduction to the Bio-Plex system including answers to the following questions:
- what is Bio-Plex technology
- why multiplex
- cost comparison of Bio-Plex to standard ELISA
- what is suspension bead array technology
- what can be analyzed with the Bio-Plex system
High resolution melt (HRM) analysis is a relatively new technique used in detecting small variations in DNA sequences between varying populations. Important applications of HRM include SNP analysis, genotyping and methylation analysis.
In the following 20 minute tutorial presented by Sean Taylor, Field Application Specialist, Bio-Rad Laboratories, you will learn the basics of high resolution melt analysis and how to practically use it in your research. The video contains information on:
- what is high resolution melt analysis, how does it work and why it is useful
- various applications for high resolution melt analysis including single nucleotide polymorphism (SNP) analysis and DNA methylation analysis
- the difference between a standard melt curved performed in routine QPCR and HRM
- the difference between non-saturation dyes such as SYBR green and saturation dyes such as evagreen
- the definition of the four class types of SNPS
- critical factors for good experimental design
- what technique you should be using for your SNP analysis (it is not one size fits all…think HRM VS Sequenome VS multiplex array
- and more!!!
For more information on HRM see A Practical Guide to High Resolution Melt Analysis Genotyping
If you are a user or potential users of Bio-Rad’s CFX or MiniOpticon systems be sure to see the message below and the accompanying real time quantitative PCR software tutorial from Bio-Rad’s Sean Taylor, Field Applications Specialist.
I am pleased to provide our recently upgraded version of CFX Manager. Version 2.0 has significant improvements and new tools from the last version 1.6 of the software. I recently did a little test drive and have recorded a 25-minute training video to help you navigate around the new features. Some key highlights include:
1. Scheduling module so that you can schedule your time on the instrument. A great tool for a multi-user instrument.
2. Master Mix Calculator…Just type in your primer concentrations and you get the volumes required for your experiment. No more thinking and more importantly…no more mistakes in volume conversions!
3. Copy and paste wells in the plate editor. I am personally very pleased with this feature.
4. Much easier access to the data you need and very nice and new data view and data export features.
The link to the video is here:
The specifications for the software can be found here:
To access the upgrade, you can go to:
Click on “Downloads” and you will find:
A. The CFX96 and CFX384 Real-Time PCR Detection Systems Firmware Update.
B. The upgrade patch for CFX Manager 2.0.
C. Precision Melt Analysis™ v1.1 Software Upgrade (This is only required for those who are using Version 1.0 of Precision Melt Analysis Software).
Important: The firmware update comes with detailed instructions that are easy to follow but do not hesitate to call tech support (1-800-4Bio-Rad) if you have any questions about the procedure.
I would like to take this opportunity to wish you and your families health and happiness in 2011!
Click here for other technical resources from Sean including his video entitled “A Practical Approach to MIQE for the Bench Scientist,” A Practical Guide to High Resolution Melt Analysis Genotyping, and the article A Simple Solution to Chromatography for High-Purity Protein Preparations: The Modular Approach
High resolution melt (HRM) analysis is a relatively new technique used in detecting small variations in DNA sequences between varying populations. Important applications of HRM include SNP analysis, genotyping and methylation analysis. The technique relies on quantitative analysis of the melt curve of a DNA fragment following amplification by PCR and in combination with qPCR permits the identification of genetic variation in nucleic acid sequences by the controlled melting of a double-stranded PCR amplicon. As opposed to standard melt curves which are run for routing qPCR experiments, HRM melt curves involve the collection of melt data in 0.2 degree C increments. Furthermore, in order to identify small nucleotide changes it is essential to eliminate background fluorescence from any HRM analysis. Recent advances in real time PCR equipment, software and reagents (including fluorescent dyes) has turned HRM into a robust analytical technique capable of detecting a small proportion of variant DNA in a background of wild-type sequence at sensitivities approaching 5%. Perhaps one of the most astounding accomplishments of HRM analysis is the ability to detect class IV SNPs (A>T or T>A mutations) which are extremely rare (they occur at a frequency of approximately 7% withing the Human Genome) and difficult to identify due to their small melt curve temperature shift.
The attached technical note from Bio-Rad Laboratories will provide you with a fantastic overview of HRM analysis and a detailed list of things to consider prior to embarking on HRM analytical experimentation. More specifically, the technical note will cover:
– important features required for HRM compatible instrumentation
– key features for HRM compatible software
-experimental design considerations for successful HRM analysis including: HRM-compatible saturating dyes, primer design and amplicon length and PCR reaction optimization
At the tech note mentions, HRM is a low-cost, readily accessible technique that can be used to rapidly analyze multiple genetic variants. Careful sample preparation and planning of experimental and assay design are crucial for robust and reproducible results. Following the attached guidelines will assist in the development of such assays.
Sean Taylor is a Field Applications Scientist at Bio-Rad Laboratories and the primary author of this tech note. Click here for other technical resources from Sean including his video entitled “A Practical Approach to MIQE for the Bench Scientist,” and the article A Simple Solution to Chromatography for High-Purity Protein Preparations: The Modular Approach