they really should be warm and shaking before transformation... try
using a cell-phone vibrator (or playstation/nintendo controller
vibrator) attached to an eppendorf tube for shaking/aeration on the
cheap
On Tue, Jan 31, 2012 at 4:07 AM, Mega <masterstorm123@gmail.com> wrote:
> 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
>> > twitter.com/onetruecathal
>> > joindiaspora.com/u/cathalgarvey
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