Working with Plasmids

Troubleshooting the plasmid DNA purification

Thi: Hi, I’m having some trouble with the yield that I’m getting after plasmid purification. I’m not sure what I might be doing wrong and was hoping if you could help me troubleshoot?

Dr. Davis: Of course! I’d be happy to help. Please describe the issues that you’re encountering and let’s try to resolve them together.

Numerous factors may contribute to low yields and poor quality of plasmid DNA. To identify the stage at which any issue may have arisen, you can check the steps outlined on Agarose Gel Analysis Of The Purification Procedure.

- What is Plasmid DNA
  - What is the main function of plasmid DNA in biotechnology?
  - Plasmid specifications
- Plasmid Purification Technologies
- DNA extraction technologies
- Working with DNA Good laboratory practice
  - Handling DNA
  - Conversions for DNA
- Working with DNA Good microbiological practice
- Isopropanol precipitation of plasmid DNA
- Storage of Plasmid DNA
- DNA cleanup
- Essential factors that may affect bacterial culture and growth
- Bacterial cultivation media and antibiotics
- Lysis of bacterial cells for plasmid purification
- Transformation of DNA
- What are endotoxins
- Troubleshooting the plasmid DNA purification process
- Quantification of DNA
- Spectrophotometric measurement of DNA concentration
  - Fluorometry
  - Agarose gel
- DNA analysis using analytical gels
- References