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electroporation

Genetics as it applies to evolution, molecular biology, and medical aspects.

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Postby JackBean » Mon Oct 24, 2011 7:41 pm

what do you mean by naked?
http://www.biolib.cz/en/main/

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Postby ZEGSUS » Tue Oct 25, 2011 12:32 pm

gDNA not into f-Plasmid
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Re:

Postby JackBean » Tue Oct 25, 2011 1:23 pm

ZEGSUS wrote: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.
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Re: Re:

Postby ZEGSUS » Tue Oct 25, 2011 3:36 pm

JackBean wrote:
ZEGSUS wrote: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?
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Postby JackBean » Tue Oct 25, 2011 3:57 pm

I really don't know, whether E. coli is capable of that. Maybe other species would be more sutable for that ;)
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Postby canalon » Wed Oct 26, 2011 3:30 am

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.
Patrick

Science has proof without any certainty. Creationists have certainty without
any proof. (Ashley Montague)
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Postby ZEGSUS » Wed Oct 26, 2011 12:16 pm

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?
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Re: electroporation

Postby ZEGSUS » Sun Nov 06, 2011 10:09 am

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?
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Postby JackBean » Sun Nov 06, 2011 10:57 am

seems just like a bunch of wires to me :-D (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?
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Re: electroporation

Postby ZEGSUS » Sun Nov 06, 2011 11:08 am

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. :evil:
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Postby JackBean » Sun Nov 06, 2011 11:25 am

I see. So even Academy is not allowed to work with GM? And I though that EU has strict rules :lol: :(
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Postby ZEGSUS » Sun Nov 06, 2011 12:42 pm

Yes.But remember -here is Bulgaria.
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