Science

On Feb 25, 2017 9:56 AM, "MC" <markyching@gmail.com> wrote:
>
> That's really interesting, I'm going to have to do some reading on recombination. Both my cultures were DH10B.

Oh, ok, I guess the-odin changed how the sell pVIB... their product page showed this week that they ship pVIB in BL21.

Hard to say what could've happened from my end, then.

I transformed it at university 4-5 years ago. Replated it on LB without Amp lots of times and it got less bright (plasmid loss), but even after 6 subcultures it was still possible to see with the naked eye. Then I left the Amp culture in the Uni fridge over the summer holidays, and after like 3 month put it on fresh LB Amp - glowed again.

So from my experience pVIB worked fine. I don't know which exact coli strain we had at Uni, they were commercial ones

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Thanks everyone!

Glass was recommended by another friend locally, and he also mentioned
looking for demolition sites.

Stainless isn't something I considered but momentarily... it indeed
would be a good choice, and I know I've seen stainless
commercial-kitchen tables with a big shelf underneath, for storage or
e.g. a microwave.


I found a piece of granite today for $5 per sq ft... there was an ad
on craigslist for free scrap pieces, I searched their free pile and
came up empty, so sent them an email asking if they had any larger
remnants or scrap, and voila!
http://imgur.com/lLjgREq

I still have to add some cross-bracing on the legs, and a shelf
underneath (I haven't decided if I want to try adding doors to the
lower area... but maybe some sliding plexiglass to keep dust out)

Thanks all!

On Fri, Feb 24, 2017 at 11:47 PM, Patrik D'haeseleer <patrikd@gmail.com> wrote:
> I'm seeing a good number of glass tabletops and stainless steel tables on
> Craigslist. With some patience, you can probably find a 2" x 4" stainless
> table for less than $100. That's not dirt cheap, but a worthwhile investment
> for something you'll be using daily for the next couple of years. Of course,
> the main drawback of stainless is that you can't use bleach on it, or it
> will rust.
>
> We actually bought a glass top table off Craigslist for CCL, just so we can
> tape some LED strip lights to it and take some CSI style lab pictures ;-)
>
> Patrik
>
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-Nathan

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When I was reading about the strain, a lot of folks complained about
recombination being possible (at least for long term maintenance of
the plasmid, or for any kind of sequence-sensitive work, like storing
a DNA library prior to sequencing)... I was actually thinking of
getting that strain and trying to select for reversions back to the
parent plasmid, using tetracycline...
https://en.wikipedia.org/wiki/PBR322

If you still have some culture around, you might try picking up some
tetracycline from a pet store and making amp+tet plates and seeing if
anything will grow.

Hard to say what happened, as I'm not too familiar with the
bioluminescent pathway and conditions for optimal light production...
but that is what comes to mind from my recent reading on pVIB and
BL21... waiting on an order myself :)

On Fri, Feb 24, 2017 at 11:24 PM, MC <markyching@gmail.com> wrote:
>> Did you grow with ampicillin?
>
>
> Yup, it came with amp media. Couldn't have been easier.
>
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Did you grow with ampicillin?

On Fri, Feb 24, 2017 at 11:01 PM, MC <markyching@gmail.com> wrote:
> My failed pVIB E. coli experiment has been bugging me and I'm wondering if
> anyone has any ideas as to why. I got the pre-transformed kit from The Odin
> which was simple enough. My initial kit did not glow out of the box so
> Josiah was kind enough to send another one which I verified was glowing. I
> made additional LB plates and streaked them using the original glowing
> plate.
>
> Easy enough right? Except none of my plates glowed. Zero. I tried different
> temps, giving them more time to grow, etc. There was also no visible
> contamination and the cultures appeared to be growing fine. What could be
> the cause of the lack of glow?
>
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On 02/23/2017 04:27 PM, Nathan McCorkle wrote:
> I'm going to scan craigslist again for some cheap granite too.

That and keep an eye open for construction cast offs.
A $20 cheap harbor freight 4 inch grinder plus a $7 diamond blade will
cut through granite when you spray water on it as you go.
Also the top edges of the cuts can be beveled that way.

Also consider remodeling trash that is "cultured marble" counter tops.
My real lab benches are made of black "cultured marble" that is dense,
fire resistant, acid proof, strong, smooth, cleanable, polishes well.
They are made of some kind of 2 part resin and rock dust in Taylor Texas.

Too bad you can't ship a 130 lb lab bench top with backsplash -- I have extra.

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I am building a small workbench to compliment my second-kitchen
countertop that is a bit smaller than I need.

I tried finding some granite or other polished rock surfaces, but they
were asking $20-$50 per square foot. I'm wondering if a finish-grade
plywood with floor polyurethane would be OK for a few months/years of
casual use.

Obviously it sucks for fire prevention (if a bunsen burner fell over,
or an alcohol lamp), as well it seems less resistant to solvents and
strong chemicals, not that I'd care if it got ugly... but it might
change the hardness and be i.e. forever sticky and messy. My main
concern is microabrasions and being harder to sterilize. I think
flooring polyurethane will probably hold up pretty well to most
equipment and labware, except maybe my metal ring-stand. I'd guess I
might see some (micro) scratching even after dragging a dusty
cardboard box across the surface, due to the rocky components of dust
(at least my garage's dust). I'd bet bleach/alcohol/lysol would be
able to combat this for only so long.

I've also considered a few pieces of large/very-large non-porous
tile... but the grout joints concern me, though I imagine I might be
able to seal them somehow and they'd be much more fire-proof, but
still would be prone to scratching since the sealant would probably be
polyurethane or epoxy.

Am I overthinking it? What have other people found
works/doesn't-work/works-for-a-while?

I'm going to scan craigslist again for some cheap granite too.

--
-Nathan

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