Home›Cell Biology›Streamlined Purification of Plasmid DNA From Prokaryotic Cultures
Cell BiologyJoVE (Open Access)Citable · DOI
Streamlined Purification of Plasmid DNA From Prokaryotic Cultures
DOI: 10.3791/2407-v
What you'll learn
✓Perform parallel clarification and plasmid DNA purification from E. coli cultures
✓Execute lysate clearance using optimized filter plates
✓Recover purified plasmid DNA and assess quality/quantity
Protocol
Biopharma Insights This protocol is a cost effective alternative for efficient parallel clarification and plasmid DNA purification from E. coli cultures. The AcroPrep Advance process starts with an optimized lysate clarification filter plate followed by purification on a high binding capacity DNA binding filter plate.
Difficulty
beginner
Total time
~2–3 hours per batch (including culture growth time separately)
Model organism
Escherichia coli
Biosafety
BSL-1
Steps
1
Clarify lysate using optimized filter plate
Load E. coli lysate onto the AcroPrep Advance clarification filter plate to remove cell debris and contaminants. This step prepares the lysate for subsequent DNA binding.
▶ 01:02
2
Bind plasmid DNA to high-capacity filter
Transfer clarified lysate to the high binding capacity DNA binding filter plate to selectively capture plasmid DNA from the cleared lysate.
▶ 02:58
3
Elute and recover purified plasmid DNA
Apply elution buffer to the DNA-bound filter to recover purified plasmid DNA in a concentrated form.
▶ 04:10
4
Quantify and assess plasmid DNA quality
Perform quantitative and qualitative analyses (spectrophotometry, gel electrophoresis, or equivalent) to verify DNA yield, purity, and integrity.