Editing Plasmid MaxiPrep HIC (section)

=== Butyl 650S to Purify SC Plasmid DNA ===

Method 1

1. Equilibrate column with 3M ammonium sulfate in Tris-EDTA buffer pH 7.4.

2. Load sample in the Tris-EDTA buffer, pH7.4 w/3 M AS to column.

3. Wash column with 2-3 column volumes of a 3M AS in TE buffer.

4. Elute DNA with a gradient of ammonium sulfate concentration from 3M to 1M over 6 CV.

the first peak contains the relaxed form of the plasmid DNA

the second peak containing the supercoiled form of plasmid DNA

Step gradient:

2.4M AS: relaxed form of the plasmid DNA<br/>1M AS: supercoiled form of plasmid DNA



'''Method 2'''

1. Dissovle sample in 50 mM Tris 10 mM EDTA pH 7.4 buffer with 2M NaCl.

2. Load to Butyl&nbsp; l650S&nbsp; 1 cm X 20 cm column with ~&nbsp; 10 ml bed volume&nbsp; equilibrated with TE containing 2M NaCl.

3. Was with TE buffer containing 2M ammonium sulfate

4. Elute DNA with TE.