Formalin-fixed, paraffin-embedded (FFPE) tissue samples are extremely common sample types. In this form, tissue is easy to store for extremely long periods of time and useful for immunohistochemical studies. Additionally FFPE samples are fairly inexpensive to produce. However the formalin fixation procedure, which was developed long before the advent of molecular biology, results in chemical crosslinking of nucleic acid and protein molecules inside the cells. This crosslinking presents a challenge for isolating intact, high-quality nucleic acid DNA; so getting at the wealth of molecular information within an FFPE sample can be difficult.
In the upcoming webinar “Successfully Overcoming the Challenges of Working with FFPE Samples”, Dr. Trista Schagat of Promega Corporation discusses some of the key considerations for anyone who is attempting to isolate nucleic acid from FFPE samples.
For instance, although the actual fixation procedure may be outside of the control of the molecular biology researcher, the process of fixation itself can have significant impact on the quality of nucleic acid isolated later on. As with any sample, the more quickly the sample arrives in the lab and the fixation process is begun, the less likely degraded nucleic acids will be a problem. Additionally, the amount of time the sample is fixed is also critical, with long fixation times creating problems in nucleic acid isolation because of increased fixation.
Before nucleic acid purification paraffin is removed from the FFPE sample using some type of organic solvent. However, organic solvents can interfere with downstream analysis if they have carried over into the purified DNA or RNA. During purification, steps are also included to reverse formalin crosslinks, remove protein, and remove contaminating nucleic acid. The success of each of these steps is highly dependent on the purification chemistry chosen. Using a purification chemistry that omits the use of organics, removes contaminants and maximizes nucleic acid recovery is important for successful downstream analyses of FFPE tissue samples.
In this webinar Dr. Schagat describes the steps involved in the purification of nucleic acids from FFPE samples, discusses the challenges, and compares different chemistries to show how they perform. She will also present data from downstream analysis (e.g., qPCR, NGS), demonstrating how the right chemistry and sample preparation can influence the quality of downstream results.
If you are interested maximizing the quality of nucleic acid from FFPE samples, register today for this free webinar to learn more.
About the Webinar Series
www.promega.com/webinars/ provides a schedule of upcoming webinars. In addition there are links to previous webinars that allow you to either view the recording or download a pdf of the presentation. There is also a pdf of additional material available for each past webinar.
To register for a webinar, use the “registration” link at: www.promega.com/webinars/ This allows you to view the webinar and participate in the live chat. Need a reminder? You can also sign-up for monthly invitations to webinars at the webinars page. Note: Live chat is only available for live webinars, not links to recorded webinars.
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