Centre for Synthetic Biology and Innovation, Imperial College London, London, United Kingdom.
Department of Bioengineering, Imperial College London, London, United Kingdom.
Sci Rep. 2016 May 31;6:26863. doi: 10.1038/srep26863.
Existing yeast genomic DNA extraction methods are not ideally suited to extensive screening of colonies by PCR, due to being too lengthy, too laborious or yielding poor quality DNA and inconsistent results. We developed the GC prep method as a solution to this problem. Yeast cells from colonies or liquid cultures are lysed by vortex mixing with glass beads and then boiled in the presence of a metal chelating resin. In around 12 minutes, multiple samples can be processed to extract high yields of genomic DNA. These preparations perform as effectively in PCR screening as DNA purified by organic solvent methods, are stable for up to 1 year at room temperature and can be used as the template for PCR amplification of fragments of at least 8 kb.
现有的酵母基因组 DNA 提取方法并不完全适合通过 PCR 对大量菌落进行广泛筛选，因为这些方法要么过于耗时费力，要么提取的 DNA 质量差，结果不一致。我们开发了 GC 预增法来解决这个问题。用玻璃珠通过涡旋混合的方法裂解来自菌落或液体培养物的酵母细胞，然后在金属螯合树脂存在的情况下煮沸。在大约 12 分钟内，可以处理多个样本以提取高产量的基因组 DNA。这些制剂在 PCR 筛选中的效果与有机溶剂法纯化的 DNA 一样有效，在室温下稳定保存长达 1 年，并且可以用作至少 8kb 片段的 PCR 扩增模板。
