How

Viewing 4 reply threads
  • Author
    Posts
    • #17593
      Helics
      Participant

      How do i insert two gene at same time inside a bacterial genome ?
      please, share any link . 🙁

    • #114617
      JackBean
      Participant

      How do you insert one gene into a bacterial genome? Why do you need to insert them at the same time? Why not consecutively?

    • #114645
      Helics
      Participant

      multiple gene insertion by conventional transformation is very time consuming and required selectable marker genes 🙁 . So I have to do different………………? Do you have any idea ????? 🙁

    • #114656
      JackBean
      Participant

      1) how do you insert single gene?

      2) what strain do you use? Is it autotroph or heterotroph?

    • #114668
      canalon
      Participant

      We would need a bit more details of what you want to do. But technically if you just want to have 2genes one after the next to insert you could either plasmids with multiple insertion sites, make your construction in those (easy) then excise the double insert and transfer it to the genomic DNA as you would do.
      If you want to be more clever you can use a super PCR (I highly recommend Phusion Taq) to build your sequence and promoters from wherever they are available and insert the product in your transfer structure.

Viewing 4 reply threads
  • You must be logged in to reply to this topic.