I have bought 'calcium chloratum' from the pharmacist. But it's not
calcium chlorate but chlorine.
I googled it up and it's used in homoeo-pathy... But is it then pure
calcium chlorine? Or has it a homeopathic dosage (1 molecule cacl2 in
1g of the salt?) ? So it would be of no sense for transformation.
I also made my own cacl2 from egg peal with HCl.... But I fear there
are proteins soluted (I should have burnt them before)
On 31 Jan., 10:25, Cathal Garvey <cathalgar...@gmail.com> wrote:
> You can get away without shaking, *if* the cultures are grown in shallow
> broth. You should still give them an occasional swirl though.
>
> And yes, they should be warm. I have successfully transformed E.coli
> grown at 30C. I incubate using a terrarium heater mat, a pet-shop
> thermostat, and a polystyrene box. I use an external thermometer to
> sanity-check the temperature, because commercial pet thermostats are
> rarely correct (but very stable, I find).
>
> If using heater wire to heat your terrarium, be careful to insulate the
> wire somehow to keep it from melting/igniting the polystyrene!
>
> On 31/01/12 09:16, Nathan McCorkle wrote:
>
>
>
>
>
>
>
>
>
> > 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 <masterstorm...@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
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>
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