Next question:
To transform bacteria they have to be in a state of exponential
growth. Is this really imperative or just increases transformation
efficiency??
____
For my transformation I think about building a breeding box using an
ATtiny, LM335, a box of polystyrol and some wire.
But in case this fails, would transformation still work?
On 27 Jan., 17:15, Mega <masterstorm...@gmail.com> wrote:
> "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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> > >>> twitter.com/onetruecathal
> > >>> joindiaspora.com/u/cathalgarvey
> > >>> PGP Public Key:http://bit.ly/CathalGKey
>
> > --www.indiebiotech.com
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