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    使用Biomek i7混合工作站自动化等离子体DNA制备可显著提高CRISPR查等高吞吐量应用的产量和质量. 这种机器人工作流提高了生产力,并减少了生成排列等离子体DNA库的错误.

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    科学领域:

    • 分子生物学分子生物学
    • 生物技术是生物技术.
    • 自动化工程自动化工程

    背景情况:

    • 高通量等离子体DNA库的准备对于CRISPR查等应用至关重要,但通常是劳动密集型和昂贵的.
    • 等离子体产量和质量的变化可能会损害整个图书馆适合下游实验.
    • 目前的手动方法容易出现人为错误,需要大量的实践时间.

    研究的目的:

    • 为高通量等离子体DNA制备和量化建立自动化工作流.
    • 为了提高生产率并减少人类干预在生成排列等离子体DNA库.
    • 为了确保下游应用的等离子体DNA的一致产量和高质量.

    主要方法:

    • 使用Biomek i7 Hybrid自动化工作站进行机器人液体处理和外围仪器集成.
    • 采用性溶解和磁珠净化用于以96孔板形式提取等离子体DNA.
    • 使用PicoGreen试验量化等离子体DNA,并将结果与NanoDrop紫外光谱光度测试进行比较.

    主要成果:

    • 自动化工作流通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验,通过PicoGreen试验.
    • 纳米滴测量显示出更高的产量 (中位数~25μg/样品,~20μg/ml培养),超过98%的样品超过10μg/ml.
    • 在PicoGreen和NanoDrop量化方法之间显示出强烈的正相关性 (R2>0.8).
    • 基因凝电泳证实了整个生成库的高质量等离子体.

    结论:

    • 开发的自动化工作流提供了一个强大的,可扩展的,和高性价比的解决方案,用于高通量阵列等离子体库的生成.
    • 自动化显著提高了等离子体DNA制备的生产率和一致性,减轻了与手工方法相关的问题.
    • 该平台非常适合要求大量高质量的等离子体DNA样本的苛刻应用.