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

🚨 Failure Case Library (1) + Submit your own case

moderate
Low DNA Yield from Gel Extraction or PCR Cleanup Due to Incomplete Elution
DNA recovery from gel extraction or PCR cleanup is lower than expected based on input amount. Some DNA likely remains bound to column.
💡 4 · ✓ 4
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