Re: Transformation questions

"Clearly, I wouldn't be so strict with DNA that was proven to have no
ecological consequences; no resistance genes, no ecological-unknowns.
A
plasmid containing only GFP and plasmid maintenance genes is hardly
worth worrying about, for example. "

I totally agree with that...

By the way, the gfp could not be taken up and expressed by plant
seemen, because the (bacterial) promotor wouldn't be readable?? It
would be junk - DNA?

On 27 Jan., 12:33, Cathal Garvey <cathalgar...@gmail.com> wrote:
> Actually, leaving the cells to "overcook" will usually destroy all
> antibiotics present; it's only unused media that really needs boiling or
> other such treatment before disposal. In the case of chloramphenicol,
> I'll probably just add some resistant cells to unused media and incubate
> in order to destroy the broth, then boil-kill and UV inactivate.
>
> Microwaves don't explicitly damage DNA.
> However, UV does, and I would advocate using UV to mince DNA before
> disposal, especially if the DNA contains medically significant (i.e.
> AmpR, maybe Chlor?) antibiotic resistance genes. Most labs have UV
> transilluminators for Gel electrophoresis, even though blue illumination
> is more all-round useful (precisely because it doesn't mince DNA).
>
> So, if I were working with E.coli, which can't survive boiling, I'd
> simply boil cellular waste in a glass beaker or flask, then put the
> container on a UV illuminator for 5 minutes before disposal.
>
> If I were working with B.subtilis, I'd autoclave rather than boiling,
> and do the same.
>
> Clearly, I wouldn't be so strict with DNA that was proven to have no
> ecological consequences; no resistance genes, no ecological-unknowns. A
> plasmid containing only GFP and plasmid maintenance genes is hardly
> worth worrying about, for example.
>
> Of course, opinions differ, and we've had animated discussions here in
> the past on this issue; whether to bother destroying antibiotics,
> whether to bother destroying DNA.
>
> On 27/01/12 08:54, Mega wrote:
>
>
>
>
>
>
>
>
>
> > Ah, you only want to dispose of the plasmids left from transformation?
>
> > On 27 Jan., 09:53, Mega <masterstorm...@gmail.com> wrote:
> >> Have you considered using UV-radiation? I saw one on a TV 'show-
> >> documentation-infotainment'  which costed about 20 bucks and they
> >> tested if it really worked.
>
> >> And, surprisingly, it really worked fine!
>
> >> Maybe Microwave sterilization could also work when it damages the DNA
> >> ( not only heat up the bacteria). Or you add e.g. ampicillin to kill
> >> the bacteria and then heat the whole pot to destroy the amp.
>
> >> On 26 Jan., 23:17, Cathal Garvey <cathalgar...@gmail.com> wrote:
>
> >>> Actually, a control Bacillus plasmid I'm going to be using is Chloramphenicol resistant: I hadn't realised it was heat resistant! I'd better imagine up a disposal procedure..
>
> >>> Venkatesh Srinivas <m...@endeavour.zapto.org> wrote:
> >>>> On Wed, Jan 25, 2012 at 11:05 PM, Tom Randall <tarand...@gmail.com>
> >>>> wrote:
> >>>>> ...
> >>>>> I am sure there are others. No environmental issues with amp or
> >>>> others
> >>>>> if you autoclave everything before you dispose of it, this will
> >>>>> inactivate most antibiotics that I am aware of. If there are any that
> >>>>> can withstand autoclaving I would be interested in knowing. This is
> >>>>> why one normally adds antibiotics to media after autoclaving the
> >>>> media
> >>>>> and cooling to ~55-60C to avoid their inactivation.
> >>>>> Go wild!
> >>>>> ...
>
> >>>> I believe chloramphenicol is thermostable; should survive autoclaving.
> >>>> Not commonly used in DIYbio I imagine though...
>
> >>>> --vs;
>
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> >>> --
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>
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