How

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Helics
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How

Post by Helics » Tue Oct 22, 2013 12:41 pm

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

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JackBean
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Post by JackBean » Wed Oct 23, 2013 7:56 am

How do you insert one gene into a bacterial genome? Why do you need to insert them at the same time? Why not consecutively?
http://www.biolib.cz/en/main/

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Helics
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Re: How

Post by Helics » Tue Oct 29, 2013 8:52 am

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 ????? :(

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JackBean
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Post by JackBean » Wed Oct 30, 2013 10:36 am

1) how do you insert single gene?

2) what strain do you use? Is it autotroph or heterotroph?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

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canalon
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Post by canalon » Sat Nov 02, 2013 6:15 pm

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.
Patrick

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