Plasmids are small, circular DNA molecules that exist in many bacteria and other organisms. They can be copied by cells independently of each other inside the same living creature which makes them an ideal way for scientists seeking new genes or therapies. Plasmid DNA is a valuable and versatile tool that can be used in many different ways, from vaccines to therapeutic drugs.
Common Challenges in Plasmid Preparation
- Plasmid Extraction Failure
The extraction time should be less than 5 minutes to avoid DNA damage. The adsorption and dissolution process needs more attention in order for plasmid cells keep their integrity during the procedure.
- Aging of E. coli
It is recommended that you reselect new colonies after coating your plate with culture.
- Low Plasmid Copies
When using a high copy number vector with the same function or increasing bacterial liquid extraction and reducing volume during elution it is important that you use an appropriate gradient.
A solution containing too few bacteria may lead to poor expression of target genes because there isn’t enough biological material for transcription- factor binding sites will not be exposed in sufficient quantities; while adding more extract might result in decreased yields since most plasmids have limited doubling capacities .
- Colonies with High False-Positive Rates, Unable to Select Correct Clones
The primers identified by colony PCR need to be designed across regions; one primer is designed on the vector, and the other primer is designed on the target fragment. This method eliminates the risk of false positives, fragments can be screened out correctly with this plasmid digesting gel and incorrect clones further confirmed by extraction.
Plasmid Preparation Applications
|Gene cloning, PCR templates, DNA markers
||Cell line construction
|Transient gene expression
||Raw materials for cell therapy
|Large-scale genome research
Plasmid Preparation | Synbio Technologies
Plasmid DNA preparation is a basic, but crucial, step in scientific research. Our plasmid preparation services provide access to unparalleled expertise and support, helping researchers confidently develop and scale up their discovery efforts. For years, Synbio Technologies has provided comprehensive, high-quality plasmid DNA preparation services to our customers from research institutes to biotechnology and biopharmaceutical companies, around the world. Our plasmid DNA preparation process is designed to meet the innovative products that are changing the world, including transfection medicines and vaccines.