DNA Purification, Quantitation and Analysis Explained

WebinarsYesterday I listened in on the Webinar “Getting the Most Out of Your DNA Analysis from Purification to Downstream Assays”, presented by Eric Vincent–a Product Manager in the Promega Genomics group.

This is the webinar for you if you have ever wondered about the relative advantages and disadvantages of the many methods available for DNA purification, quantitation and analysis, or if you are comparing options for low- to high-throughput DNA purification. Eric presents a clear analyses of each of the steps in a basic DNA workflow: Purification, Quantitation, Quality Determination, and Downstream Analysis, providing key considerations and detailing the potential limitations of the methods commonly used at each step.

The DNA purification method chosen has an affect on the quality and integrity of the DNA isolated, and can therefore affect performance in downstream assays. Accuracy of quantitation also affects success, and the various downstream assays themselves (such as end-point PCR, qPCR, and sequencing) each have different sensitivities to factors such as DNA yield, quality, and integrity, and the presence of inhibitors.

Purification: Setting the Stage for Success
The DNA purification method that you choose will be influenced by factors such as sample volume and throughput requirements, productivity and cost issues, as well as the stringency of the requirements of your downstream assay. The webinar covers many DNA purification options: low cost manual precipitation-based systems like the Wizard Genomic System; small-scale automated systems, which increase throughput and give more consistent sample preparation but involve a higher cost investment; manual 96-well plate-based methods using silica membranes, which are good low-cost options for labs that process large numbers of samples infrequently; and finally large-scale, fully-automated high-throughput systems that can integrate both sample prep and downstream assays, bringing huge benefits in productivity and consistent processing, but requiring a much larger cost investment.

As well as discussing the various throughput options affecting DNA purification choices, the webinar also highlights the problems of DNA purification from difficult sample types such as FFPE tissues, which may be months or years old and have DNA damage caused by fixation. Sample data generated using the ReliaPrep FFPE system is presented to highlight some of the issues encountered with this common sample type, and to illustrate methods for assessing DNA quality.

Quantitation: How Do All These Methods Work?
Moving on to DNA Quantitation Systems, the webinar provides a useful summary of UV Spectroscopy (focusing on the NanoDrop Spectrophotometer), Fluorescent DNA binding dyes, Gel Electrophoresis, and the Agilent Bioanalyzer. Listen to the webinar to find out how each of these systems work, how easy they are to use, what their strengths are, and to learn the potential disadvantages of each method.

Downstream Assays: How does DNA Quality Affect My Choices?
Finally, various downstream assays are discussed, also focusing on DNA quality, quantity and purity requirements, and covering considerations for choosing each particular assay type.

Watch the Webinar!
The webinar recording is now available online here. It’s a great introduction to the methods available for DNA purification, quantitation and analysis for anyone who wants to know a little more about the basis of each method and the considerations to bear in mind to identify the best systems for their needs.

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Isobel Maciver

Isobel is a graduate of the University of Edinburgh and of Aston University in Birmingham, U.K. She is a technical writer and editor, and is also manager of the Scientific Communications group at Promega. She enjoys writing about issues in science and communication.

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