Wright W E
Department of Cell Biology and Neuroscience, University of Texas Southwestern Medical Center, Dallas 75235.
Biotechniques. 1992 Nov;13(5):772-9.
A technique called partial-digest sequencing that permits DNA of 4-6 kb in length to be sequenced without subcloning is described. The method exploits the specific cuts introduced by partial digestion with restriction endonucleases that have 4-base recognition sites to produce ordered ladders of PCR-amplified fragments. The staggered ends contain PCR primers and can thus be individually sequenced using conventional methods to yield overlapping sequences covering the entire region. This method should have significant impact on both large and small DNA sequencing projects and find many applications in general manipulations in which ordered sets of deletions need to be produced.
本文描述了一种名为部分消化测序的技术，该技术可对长度为4 - 6 kb的DNA进行测序而无需亚克隆。该方法利用具有4个碱基识别位点的限制性内切核酸酶进行部分消化所引入的特异性切割，以产生PCR扩增片段的有序梯状条带。交错末端含有PCR引物，因此可以使用常规方法单独测序，以产生覆盖整个区域的重叠序列。该方法对大型和小型DNA测序项目都应具有重大影响，并将在需要产生有序缺失集的一般操作中有许多应用。
