本文介绍利用固相法来进行限制性酶切位点的绘制,按照作者的方法,可以避免在得到单一末端标记DNA过程中的复杂工作,并且不需要使用放射性物质。作者在PCR反应中使用一个末端带有生物素标记的引物和一个不带有标记的引物来扩增所需的DNA片段。这些引物可存在于DNA片断中或载体侧翼的已知顺序中。得到的线状DNA将在一端带有生物素标记。然后线状DNA被连接在带有链霉亲合素的磁珠上,通过洗脱去掉杂质,再选择合适的限制性酶直接对结合在磁珠上的DNA进行部分酶切。上述方法与已建立的固相测序法相似。除了标上 This article describes the use of solid-phase method for restriction enzyme sites, according to the author’s method, to avoid the complex work of a single end-labeled DNA, and does not require the use of radioactive material. The authors used a primer with a biotin end and a primer without a primer to amplify the desired DNA fragment in a PCR reaction. These primers may be present in the DNA sequence or in a known sequence flanking the vector. The resulting linear DNA will have a biotin label on one end. The linear DNA is then ligated to streptavidin-coated magnetic beads, the impurities are removed by elution, and the DNA bound to the magnetic beads is partially digested directly by selecting appropriate restriction enzymes. The above method is similar to established solid-phase sequencing. Except marked