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June 01, 2011 (Vol. 31, No. 11)

HT Purification of Cell and Tissue DNA

High-Throughput Workflow Solution Now Available to Obtain High Yields of Pure DNA

  • Results

    Click Image To Enlarge +
    Figure 1. Simultaneous purification was performed from mouse ear, liver, and kidney samples and high levels of DNA were obtained.

    The data obtained from the purification of various cell and tissue samples demonstrated that high yields of quality DNA are easily extracted with the KingFisher Cell and Tissue DNA Kit.

    DNA purified from mouse ear, liver, and kidney samples produced a high yield of DNA, as shown in Figure 1. This is demonstrative of the ability to provide quality starting material for applications, such as the genotyping of mice, where ear and tail are typically used. DNA yields obtained depend on both the lysis time and volume of tissue; 10 mg and 15 mg of mouse kidney samples were lysed for 1 hour, 4.5 hours, and overnight, the results of which are shown in Figure 2.

  • Click Image To Enlarge +
    Figure 2. 10 and 15 mg samples of mouse kidney were lysed for 1 hour, 4.5 hours, or overnight, followed by DNA purification.

    However, even after only 1 hour of lysis, the DNA obtained was of an excellent yield and ratio, making it ideal for use in downstream applications. In addition, lysis of the ear, liver, and kidney samples in the KingFisher Flex for 15 minutes was effective, producing an adequate yield for subsequent PCR reactions (data not shown).

  • Click Image To Enlarge +
    Figure 3. Dilution series from HeLa-S3 cells indicating that the purified DNA yield correlated with the quantity of cells within the sample.

    The effects of the amount of sample material were compared with the purification of DNA from a different quantity of HeLa-S3 cells. The results indicate that there is a direct correlation between the yield of purified DNA samples and the number of cells in the sample, as shown in Figure 3. Even with the smallest amount of approximately 15,000 cells, the purification process was efficient and the subsequent PCR was successful.

  • Conclusion

    Efficient and fast DNA purification is an essential starting point to a broad array of drug discovery applications. As such, it is essential that researchers have a reliable method of obtaining high yields of pure DNA. The KingFisher Cell and Tissue DNA Kit, in combination with the KingFisher Flex offers a high-throughput workflow solution.

    Since different sample materials require a range of lysis times, the entire purification process can be effectively optimized to take into account the volume and type of material being used. As a result, optimal yields are produced from each purification for use in downstream applications. Additionally, with some materials, the hands-on time can be minimized and throughput maximized by performing the lysis directly in the KingFisher instrument.

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