Team:Paris/Miniprep

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Adaptation of PureYield(TM) Plasmid Miniprep System (Promega)

Prepare Lysate

• Add 2ml of bacterial culture to a 2ml microcentrifuge tube.
• Centrifuge 1.5ml of bacterial culture for 30 seconds at maximum speed in a microcentrifuge. Discard the supernatant.
• Add 600µl of H₂O (Gibco), and resuspend completely.
• Add 100µl of Cell Lysis Buffer (Blue), and mix by inverting the tube 6 times.
• Wait 2min but no more than 5min
• Add 350µl of cold (4-8°C) Neutralization Solution, and mix thoroughly by inverting.
• Centrifuge at maximum speed in a microcentrifuge for 10 minutes.
• Place a PureYield(TM) minicolumn on a Luer-Lok(R) adapter of a VacMan(R).
• Transfer the supernatant (~900µl) into a PureYield(TM) minicolumn without disturbing the cell debris pellet (yellow-orange).
• Apply vacuum pulling the lysate through the column.

Wash