Is a 5` phosphate needed for homologous recombination?
I am a bit confused whether phosphorylated DNA is needed to achieve efficient homologous recombination in e coli. I believe there are methods that use ssDNA oligos or PCR products as inserts for recombineering so these would usually not have a 5` phosphate but there also seems to be some literature that makes it sound like it is beneficial.
Can anyone clarify this, please.
Thanks
Hi Moein,
Usually, bacteria have RecA that mediate homologous recombination. so we don'd need 5'phosphate but if you are performing after DNA ligation (in case of cloning), the ligase enzyme we use have 5' phosphate and 3' hydroxyl for nick sealing. So it depends on your experimental stage. Bottomline is, it's not strictly required but can be beneficial if used. Hope it helps. Good luck!
- Badges
- Science topic
- Similar topics
- Medicine
- Disease
- Infectious Diseases