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Best Practices for Handling Genomic DNA During NGS Library Preparation
EP34589
Poster Title: Best Practices for Handling Genomic DNA During NGS Library Preparation
Submitted on 16 Dec 2020
Author(s): Chava Pocernich, Kyle Luttgeharm, and Solange Borg
Affiliations: Agilent Technologies
This poster was presented at ASHG 2019
Poster Views: 455
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Poster Information
Abstract: Superior NGS library preparation and sequencing results start with quality genomic DNA (gDNA). Quality control analysis provides a great deal of information that is required before beginning NGS library preparation including size, smear distribution, and concentration. Quality of the gDNA starting material can be modified by the dynamic environment and handling processes it encounters. To determine how common laboratory practices such as pipetting, vortexing, and freeze-thaw cycling impact gDNA quality, commercially available gDNA samples were analyzed using the Agilent 5200 Fragment Analyzer system. Both the Agilent Genomic DNA 50 kb kit and HS Genomic DNA 50 kb kit were used to determine the average size, smear distribution pattern, concentration, and percent of degradation of the gDNA sample. It was found that different sample handling practices have varied impact on gDNA sizing, with some, such as high-speed vortexing having a greater effect on sample integrity compared to gentler handling practices, such as slow pipetting with a wide-boar pipet tip. From this study, we have developed a set of best practices for handling gDNA. Quality control analysis of gDNA and gentle handling practices ensure gDNA integrity and build confidence in the success of downstream applications such as NGS library preparation.
For Research Use Only. Not for use in diagnostic procedures.
Summary: An overview of the gDNA best handling practices during NGS library preparationReport abuse »
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