Principle

Buffer A3 neutralizes the resulting lysate and creates appropriate conditions for binding of plasmid DNA to the silica membrane of for example the NucleoSpin® Plasmid/ Plasmid (NoLid) or NucleoSpin® Plasmid QuickPure Column. Precipitated protein, genomic DNA, and cell debris are then pelleted by a centrifugation step. The supernatant is loaded onto a NucleoSpin® Plasmid/Plasmid (NoLid) or NucleoSpin® Plasmid QuickPure Column.

Lysate neutralization and LyseControl

Proper mixing of the lysate with Neutralization Buffer A3 is of utmost importance for complete precipitation of SDS, protein, and genomic DNA. Incomplete neutralization leads to reduced yield. However, released plasmid DNA is very vulnerable at this point and shaking too much or too strongly will damage the DNA.