Editing Qiagen MaxiPrep (section)

== Anion-Exchange Chromatography using Qiagen MaxiPrep Column<br> ==

1. Resuspend E. coli cells in XX mL Buffer P1.

<br>2. Add xx mL Buffer P2, mix the solution, and incubate at room temperature for 5 min.<br>

3. Add xx mL Buffer P3 and mix carefully.<br>

4. Incubate the lysate for 30 min at room temperature (15â25 oC) to allow the flaky white precipitate of SDS, protein, genomic DNA, and cell residue to rise to the surface.<br>

5. Carefully pump the lysate out of the bottle.<br>

6. Mix the lysate with 0.10 vol of buffer ER (QIAGEN) and filter the mixture through a QIAfilter unit (QIAGEN). Collect the filtrate for subsequent chromatography.

<br>7. Equilibrate the QIAGEN Ultrapure 100 Column with 350 mL buffer QBT.<br>

<br>8. Load the column with the filtrate from step 6.

<br>9. Wash the charged column with&nbsp; Buffer QC.

<br>10. Elute plasmid DNA with&nbsp; Buffer QN.

<br>11. Precipitate the DNA with 0.7 vol of isopropanol at 4 oC and centrifuge at 6000 rpm for 10 min at 4 oC.

<br>12. Wash the DNA pellet with&nbsp; 75% ethanol.

<br>13. Dry the DNA pellet and resuspend it in the appropriate buffer for further applications.<br>

[[Category:Protocols]]