Moreira D
Laboratoire de Biologie Cellulaire BC4, URA CNRS 2227, Université Paris-Sud, Bâtiment 444, 91405 Orsay Cedex, France.
Nucleic Acids Res. 1998 Jul 1;26(13):3309-10. doi: 10.1093/nar/26.13.3309.
The use of agarose blocks containing embedded DNA improves the PCR amplification from templates naturally contaminated with polysaccharides or humic acids, two powerful PCR inhibitors. Presumably, the difference in size between the DNA macromolecules and these contaminants allows their effective removal from the agarose blocks by diffusion during the washing steps, whereas genomic DNA remains trapped within them. In addition, agarose-embedded DNA can be directly used for PCR since low melting point agarose does not interfere with the reaction. This simple and inexpensive method is also convenient for genomic DNAs extracted by other procedures, and it is potentially useful for samples containing other kinds of soluble inhibitors, overcoming this important problem of current amplification techniques.
使用含有嵌入DNA的琼脂糖块可改善从天然被多糖或腐殖酸污染的模板进行的PCR扩增,多糖和腐殖酸是两种强大的PCR抑制剂。据推测,DNA大分子与这些污染物之间的大小差异使得它们在洗涤步骤中通过扩散从琼脂糖块中有效去除,而基因组DNA则保留在其中。此外,由于低熔点琼脂糖不干扰反应,因此嵌入琼脂糖的DNA可直接用于PCR。这种简单且廉价的方法对于通过其他程序提取的基因组DNA也很方便,并且对于含有其他种类可溶性抑制剂的样品可能有用,克服了当前扩增技术的这一重要问题。