GC Preps: Fast and Easy Extraction of Stable Yeast Genomic DNA.
GC Preps: Fast and Easy Extraction of Stable Yeast Genomic DNA.
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DOI：
10.1038/srep26863
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发表时间：
2016-05-31
影响因子：
4.6
通讯作者：
Ellis T
中科院分区：
文献类型：
--
作者：
Blount BA;Driessen MR;Ellis T
现有的酵母基因组DNA提取方法由于太长、太费力或产生质量差的DNA和不一致的结果而不理想地适合于通过PCR广泛筛选菌落。我们开发了GC制备方法来解决这个问题。通过与玻璃珠涡旋混合来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下煮沸。在大约12分钟内，可以处理多个样品以提取高产量的基因组DNA。这些制备物在PCR筛选中与通过有机溶剂方法纯化的DNA一样有效，在室温下稳定长达1年，并且可以用作至少8kb片段的PCR扩增的模板。
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.