Hicks Linda, van der Graaf Charlotte M, Childress Jacob, Cook Emily, Schmidt Karen, Rosenzweig Frank, Kroll Eugene
Division of Biological Sciences, University of Montana, Missoula, MT 59812, USA.
J Biol Methods. 2018 Mar 9;5(1):e86. doi: 10.14440/jbm.2018.218. eCollection 2018.
Genome analysis using pulsed-field gel electrophoresis (PFGE) has been used in applications ranging from typing bacterial strains to radiobiology to cancer research. While methods for running PFGE have been significantly improved since its invention, the method for preparing chromosomal DNA itself has remained essentially unchanged. This limits the applicability of PFGE, especially when analyses require many samples. We have streamlined sample preparation for routine applications of PFGE through the use of deep-well 48-well plates. Besides saving time, our protocol has the added advantage of reducing the volume of expensive reagents. Our improved protocol enables us to reduce throughput time and simplify the procedure, facilitating wider application of PFGE-based analyses in the laboratory.
使用脉冲场凝胶电泳（PFGE）进行的基因组分析已应用于从细菌菌株分型到放射生物学再到癌症研究等诸多领域。自PFGE发明以来，其电泳运行方法已有显著改进，但染色体DNA本身的制备方法基本未变。这限制了PFGE的适用性，尤其是在分析需要处理大量样本时。我们通过使用48孔深孔板简化了PFGE常规应用的样本制备过程。除了节省时间外，我们的方案还有减少昂贵试剂用量的额外优势。我们改进后的方案能够缩短通量时间并简化操作流程，有助于在实验室更广泛地应用基于PFGE的分析方法。
