Figure 3.

DNA extraction with CTAB in the presence of Co³⁺(NH₃)₆Cl^–₃. (A) Outline of the protocol. Experimental details are given in Materials and Methods. (B) Branch migration during yeast DNA extraction using the protocol. R1 and R2 were annealed in TNM buffer. As a positive control for branch migration, an aliquot was incubated at 37°C in TNM buffer (10 mM MgCl₂) for the indicated times. Remaining annealed substrate was added to yeast spheroplasts, and DNA was extracted as described in Materials and Methods. A sample was taken after chloroform:isoamyl alcohol extraction (CHCl₃). DNA pellets obtained after ethanol precipitation from NaCoHex solution (NaCoHex) and after ethanol precipitation from TMNa solution (TMNa) were resuspended in TMSpe buffer on ice. Branch migration was measured as described in Figure 2; net product refers to the fraction of total ³²P label migrating as monomer duplex, after subtraction of residual unannealed substrate.