Path: utzoo!attcan!uunet!husc6!linus!philabs!aecom!diaz From: diaz@aecom.YU.EDU (Dizzy Dan) Newsgroups: sci.bio Subject: Obtaining single-stranded linear DNA Message-ID: <1792@aecom.YU.EDU> Date: 11 May 88 03:49:37 GMT Organization: Graduate School of Hard Knocks Lines: 16 Here's this week's biochemical puzzler (you all did well on the antisense RNA trick question): I am interested in obtaining tritiated (or otherwise labeled) single-stranded linear DNA of uniform length (has to be at least 1knt long) for exonuclease assays. M13 seems ideal since the cells will spit out labeled ssDNA (circular) if we cook them with label. How do we make the DNA linear? Do we shear it, or cut it with a restriction endonuclease with ss activity (expensive!) or what? The DNA does not have to be linearized in the same place, but it must be linear, and cut once, or at most a few times. -- dn/dx Dept Molecular Biology diaz@aecom.yu.edu Dizzy Dan Al Einstein's Med School Big Bad Bronx, NY