Assessment of Tissue Lysis Methods for DNA Extraction from Murine Spleen
Dahlberg, Kaylin
Dahlberg, Kaylin
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Abstract
For a number of laboratories, DNA extractions from tissue are the first step for a PCR protocol. Portions of tissue are lysed, or broken down from solid tissue into liquid culture using a myriad of reagents and techniques. Many laboratories, including ours, employ a bead beater protocol to lyse a small segment of tissue. Since we utilize these extractions for measuring viral load titer, we questioned whether using the bead beater and a 10-�g sample was representative of the entire tissue. Another common lysing technique is complete homogenization of tissue using sonication. Sonication is the act of using sound waves in order to break tissue. We hypothesized that the viral load titer of the samples that were sonicated will not be significantly different than those that were broken down using the bead beater. With this experiment, we utilized spleen from mice strains that were 9 days post infection with Murine Gammaherpesvirus-68 (MHV-68). DNA was isolated from the same third of the spleen for each method; with 10 �g being isolated prior to sonication of the rest of the sample. The extraction protocol after tissue breakdown followed the protocol identified in the Qiagen DNeasy kit. The success of DNA extraction was measured using a qPCR, with DNA normalized to 35�g/�L. From this, it was determined that the method of extraction; either sonication or utilization of a bead beater, did not have a significant impact on the amount of viral DNA measured. This finding allows for continuation of protocols that have been utilized within the laboratory and ensures that there will be tissue samples left over after DNA isolation for future experiments. There is not much literature exploring the comparison between using a bead beater or sonication for DNA extraction.
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Date
1/1/2026
Student Status
Junior (Graduating in 2027)
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Poster
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Biochemistry and Medical Diagnostics
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College of Arts and Sciences
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Clinical Science