electroporation

[JackBean]

what do you mean by naked?

[ZEGSUS]

gDNA not into f-Plasmid

[JackBean]

http://aem.asm.org/cgi/reprint/60/11/4192.pdf
Well I guess that is possible to insert "naked" DNA into bacteria.

yeah, but I guess it depends on the species. Agrobacterium is pathogenic bacteria and some of its relatives (I don't know whether Agrobacterium by itself) can posses linear plasmids. Such bacteria will of course accept "naked" DNA (but still the problem with replication), but it's questionable how will E.coli do.

[ZEGSUS]

http://aem.asm.org/cgi/reprint/60/11/4192.pdf
Well I guess that is possible to insert "naked" DNA into bacteria.

yeah, but I guess it depends on the species. Agrobacterium is pathogenic bacteria and some of its relatives (I don't know whether Agrobacterium by itself) can posses linear plasmids. Such bacteria will of course accept "naked" DNA (but still the problem with replication), but it's questionable how will E.coli do.

You mean that the linear DNA cant be save for the future generations?

[JackBean]

I really don't know, whether E. coli is capable of that. Maybe other species would be more sutable for that

[canalon]

E. coli will not readily take up naked DNA. Other species can, but not E. coli.
If you are looking for a specific gene from Acetobacter bulgaricus your best bet is indeed to try a BAC. There are many kits commercially available (Google "BAC cloning kit" and your country of residence to find some supplier, or ask your friendly local sales rep) to do just that.
And no, not all bacteria will accept DNA by electroporation. But in your case that should not be a problem.

[ZEGSUS]

In my country the gene engineering is forbidden so there is no suppliers here.And do you have a easy protocol how to insert gDNA into E.coli?

[ZEGSUS]

I want to report a little success about my project in electroporation.This is my first prototype of electroporator.I have succeeded to make it with less than 100$.What do you thing?

[JackBean]

seems just like a bunch of wires to me (I don't have much big screen here). Just curious, where are you from, that gene engineering is forbidden? You mean even manipulation with bacteria, right?

[ZEGSUS]

http://en.wikipedia.org/wiki/Bulgaria
Yes even manipulation with bacteria. The "eco organisation" provides a lot of pressure on the government. And the government ban GMO.

[JackBean]

I see. So even Academy is not allowed to work with GM? And I though that EU has strict rules

[ZEGSUS]

Yes.But remember -here is Bulgaria.