Chai J
Institute of Genetics, Fudan University, Shanghai.
Yi Chuan Xue Bao. 1990;17(2):136-42.
A procedure for rapid restriction mapping of cosmid or lambda phage clones has been developed. The mapping of cosmid is based on linearization of circular cosmid DNA in vitro by the phage lambda terminase. Partial digestion products are selectively labelled at the right or left cos cohesive termini by hybridization with [32P] oligonucleotides complementary to the single-strand cos end. After gel electrophoresis and autoradiography, the restriction map can be directly determined from the "ladder" of partial digestion products of cosmid or lambda clones with computer program or by hand.
已开发出一种用于黏粒或λ噬菌体克隆快速限制酶切图谱分析的方法。黏粒的图谱分析基于λ噬菌体末端酶在体外使环状黏粒DNA线性化。通过与与单链cos末端互补的[32P]寡核苷酸杂交,部分消化产物在右侧或左侧cos黏性末端被选择性标记。经过凝胶电泳和放射自显影后,可通过计算机程序或手工从黏粒或λ克隆的部分消化产物“阶梯”直接确定限制酶切图谱。