I need to send Fosmid DNA for sequencing, but they told me the concentrations were too low & there are multiple bands. I tried mini-prep Qiagen kit many times & also midi-prep. How can I make sure the multiple bands are not degradation or genomic DNA?
Hi Laila,
Is the fosmid in uncut state in your picture? Did you attempt to digest your fosmid by restriction enzyme with single cutting site?
In my experience, DNA degradation will reveal smear form on gel.
The genomic DNA should be aggregated and removed in the mini or midi-prep procedure. Check the pH value of denature and renature buffer.
Good luck.
Weber
Hello Laila,
Sometimes, the multiple bands represent the supercoiled and non-supercoiled forms of plasmids/fosmids. So, I agree with Weber; you should digest your fosmid with a suitable restriction enzyme. If it is completely digested, you should see only one band of linearized DNA if your fosmid is pure.
Good luck Laila!
- Badges
- Science method