Bio-Rad Quantum Prep Plasmid Kits - Manual (Page 8)

Tris-HCl, 0.1 mM EDTA, pH 8.0). Eluting with H
2
O or TE
preheated to 70°C can also improve the yield. However, these
high temperatures may partially denature large plasmids.
• To increase the concentration of DNA eluted, use one half the
recommended volume of H
2
O or TE (50 µl). Alternatively, the
purified DNA may be ethanol-precipitated and resuspended in
10–20 µl of water or TE to achieve a higher concentration. See
Figure 1 for a detailed comparison of DNA yield and
concentration versus elution volume.
2.2 Protocol
All centrifugation steps are performed at maximum speed
(12,000–14,000 x g).
1. Transfer an overnight culture (1–2 ml) of plasmid-containing
cells to a microcentrifuge tube. Pellet the cells by centrifugation
for 15–30 secs. Remove all of the supernatant by aspirating or
pipeting.
2. Add 200 µl of the cell resuspension solution and vortex or pipet
up and down until the cell pellet is completely resuspended.
3. Add 250 µl of the cell lysis solution and mix by gently inverting
the capped tube about ten times (do not vortex). The solution
4