I extract plasmid DNA by cell lysis extraction method but the yield is so low in contrast with some commercial kits like Quigene or GeneAll. What are possible reasons?
Multiple factors could be involved such as poor culturing conditions, preparation of plasmid and insufficient or excessive amounts of starting materials. You should be optimized for all different factors.
The vector can be a low copy number which gives very few copy per bacteria. Other factors depending of your protocol is trapping of the Plasmid DNA in the protein and Genomic DNA precipitation. You should maybe explain your protocol from there we could give you some possible tracks of improvement
Dear Dehghani,
There must be some problem in lysis and denaturation buffer due to which cell doesn't lysed properly and plasmid doesn't come out of the cell. And if it come out accurately then your lysis buffer doesn't work efficiently means it under or over lysed denature the DNA
It is depending on several factors such as the bacterial culture and its growing conditions, type of plasmid, or experimental errors.
You should consider and optimize all these factors.
Hope this helps
- Badges
- Science topic