Home › Cell Biology › PureYield™ Plasmid Miniprep System
Steps
  1. 1 Prepare bacterial culture and lyse cells 00:23
  2. 2 Neutralize lysate with neutralization solution 00:38
  3. 3 Centrifuge lysate and assemble column 01:18
  4. 4 Load supernatant onto mini column 01:42
  5. 5 Wash column with endotoxin removal solution 02:08
  6. 6 Perform final column wash step 02:37
  7. 7 Elute plasmid DNA from column 02:59
Cell Biology Promega Corporation

PureYield™ Plasmid Miniprep System

Protocol
Difficulty
intermediate

Steps

1
Prepare bacterial culture and lyse cells

Add 100 microliters of blue cell lysis buffer to 600 microliters of bacterial culture and mix by inverting to lyse the cells.

▶ 00:23
2
Neutralize lysate with neutralization solution

Add 350 microliters of cold neutralization solution to the lysate and mix by inverting until the color changes completely from blue to yellow.

▶ 00:38
3
Centrifuge lysate and assemble column

Centrifuge the lysate at maximum speed in a microcentrifuge for 3 minutes while simultaneously assembling the PureYield mini column.

▶ 01:18
4
Load supernatant onto mini column

Transfer the supernatant to the PureYield mini column and centrifuge at maximum speed for 15 seconds to bind the plasmid DNA.

▶ 01:42
5
Wash column with endotoxin removal solution

Add 200 microliters of endotoxin removal wash to the mini column and centrifuge at maximum speed for 15 seconds to remove contaminants.

▶ 02:08
6
Perform final column wash step

Add 400 microliters of column wash solution to the mini column and centrifuge at maximum speed for 30 seconds to thoroughly clean the column.

▶ 02:37
7
Elute plasmid DNA from column

Transfer the mini column to a clean 1.5 milliliter microcentrifuge tube, add 30 microliters of elution buffer directly to the column matrix, let stand for 1 minute at room temperature, then centrifuge at maximum speed for 15 seconds to elute the plasmid DNA.

▶ 02:59
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