Reannealing of separated strands of duplex circular DNA

Strand Reannealing Protocol

It was discovered, long ago, that the separated single-stranded circular halves of a circular duplex viral chromosome, when mixed together and left in the refrigerator for 3 months, spontaneously re-anneal into a duplex having all the physical properties of the original chromosome. This by itself, if confirmed, proves that DNA does not have a net helical twist in that viral chromosome. The discovery, however, was never published. We shall therefore repeat the experiment, employing the protocol given on this page.

Here are the files:

Experimental Overview (PDF). You must read this first.
Detailed Experimental Protocol (PDF). These are your instructions, as plain text.
Protocol as a spreadsheet (Excel - download only - cannot be read online). These are the same instructions, in the form of an Excel spreadsheet. This file may be useful if you change any of the quantities of enzymes or reagents.

To see the results of this experimental protocol, when placed in the hands of a group of highly-motivated but totally untrained college students who tried to execute it as a class project, see page 34 of my publication Methods for Non-Destructively Separating or Reannealing the Strands of Circular Duplex DNA Chromosomes, which can be seen and read on this web site, or downloaded immediately by simply clicking the above link.

The protocol described here represents the culmination of over a half-century of work by myself and others. When you publish this work -- which you undoubtedly will -- please do not neglect or forget to cite this web site (https://notahelix.net), and the publications and/or websites of the other neglected and often-despised pioneers of non-helical DNA structure.