Ishihara H, Shikita M
Division of Chemical Pharmacology, National Institute of Radiological Science, Chiba, Japan.
Anal Biochem. 1990 Feb 1;184(2):207-12. doi: 10.1016/0003-2697(90)90670-5.
A rapid, simple, and efficient method for DNA blotting is presented. This method is characteristic in that DNA is transferred without denaturation from an electrophoretic gel to a membrane in low-salt concentrations and denatured on the membrane after blotting. More than 89% of double-stranded DNAs ranging in size from 75 to 1.9 million bp can simultaneously be transferred from the gel to a positively charged nylon membrane within 10 min. The present "low-salt electroblotting" method is superior to other blotting methods in that it saves time and labor and its high and even transfer efficiency makes it useful for hybridization analysis.
本文介绍了一种快速、简单且高效的DNA印迹法。该方法的特点是DNA在低盐浓度下从电泳凝胶转移至膜上时无需变性,而是在印迹后在膜上进行变性。大小在75至190万碱基对之间的双链DNA中,超过89%能够在10分钟内同时从凝胶转移至带正电荷的尼龙膜上。目前的“低盐电印迹”方法在节省时间和人力方面优于其他印迹方法,其高且均匀的转移效率使其适用于杂交分析。