Reynolds T R, Uliana S R, Floeter-Winter L M, Buck G A
Department of Microbiology and Immunology, Medical College of Virginia Campus, Virginia Commonwealth Univ. Richmond 23298.
Biotechniques. 1993 Sep;15(3):462-4, 466-7.
We describe optimization of a coupled amplification and cycle sequencing (CAS) method for rapid characterization of cloned or genomic DNA. Our modification of this method, termed coupled PCR amplification and cycle sequencing (CPACS), utilizes commercially available reagents, does not require template purification and produces high-quality sequence ladders from nanogram quantities of complex genomic DNA. The reactions have been streamlined to permit automation. Finally, we show that the technique can be applied more efficiently in conjunction with the AutoTrans 350 Direct Transfer Electrophoresis System and 33P-labeled sequencing primers.
我们描述了一种用于快速鉴定克隆DNA或基因组DNA的耦合扩增与循环测序（CAS）方法的优化。我们对该方法的改进，即耦合PCR扩增与循环测序（CPACS），使用市售试剂，无需模板纯化，并且能够从纳克量的复杂基因组DNA中产生高质量的序列梯。反应流程已简化以实现自动化。最后，我们表明该技术与AutoTrans 350直接转移电泳系统和33P标记的测序引物结合使用时效率更高。
