Quick Denature Plasmid Sequencing Kit

[Colin Rasmussen]

We do an earlier version of the NaOH denaturation technique.  With 20ul 
of mini-prep DNA we add 5ul of denaturing solution (39ul water, 10ul 10N 
NaOH, 1ul 0.5M EDTA), sit 10 min at room temperature, add 3ul 3M NaOAc, 
80ul 95% EtOH and collect the DNA by centrifugation (10 min at 12 kxg).  
We then add follow the standard Sequeunase 2.0 protocol.  Never fails.