Purification of plasmid DNA on a UNOsphere Q column. Clarified bacterial lysate (10 ml, adjusted to pH 8.0) was loaded onto a 0.5 x 11 cm column (2.1 ml) in buffer A (10 mM sodium phosphate, 0.3 M NaCl, 1 mM EDTA, pH 8.0). The sample was eluted with a 0–40% gradient of buffer B (10 mM sodium phosphate, 1.0 M NaCl, 1 mM EDTA, pH 8.0) at a flow rate of 2 ml/min (600 cm/hr). The column was washed with 10 column volumes of 40% buffer B, followed by 100% buffer B for 5 column volumes. The effluent was monitored at 254 nm. Each fraction was 5 ml. Blue, A254; red, conductivity; black, theoretical gradient.
Analysis of plasmid DNA purified on UNOsphere Q support. Fractions were separated on a 0.8% agarose gel. Lane 1, 1 kb marker (catalog #170-8204); lane 2, crude lysate; lane 3, flowthrough (fractions 2–6); lane 4, fractions 9–18; lane 5, fractions 19–30; lane 6, fractions 31–35; lane 7, fraction 36.
Restriction enzyme analysis of plasmid DNA purified on UNOsphere Q support. Lane 1, 1 kb marker; lane 2, undigested clarified lysate; lane 3, EcoRI-digested clarified lysate; lane 4, undigested fractions 31–35; lane 5, digested fractions 31–35; lane 6, undigested fraction 36; lane 7, digested fraction 36.
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