Re: Closet biologists story

From: Eugene Leitl (Eugene.Leitl@lrz.uni-muenchen.de)
Date: Wed Jan 23 2002 - 12:00:30 MST


On 24 Jan 2002, Alejandro Dubrovsky wrote:

> > a) Water purification: $3000-5000
> ouch. maybe buy the end product instead.

I think a used Millipore can be cheaper, especially if you yourself do the
maintenance. If you can limit yourself to small quantities, a constantly
running distiller (a glass shenanigan with a quartz heating rod) could be
substituted. You'd still want to use a sterile filter at the tap end. And,
think about electricity bill, and a trickle of an output. I would
recommend calling your local Millipore rep, and asking him for a quote on
a used setup.

> > b) -80 deg freezer: $3000-5000
> is this definitely necessary? -15C won't do?

Nope. In fact, sometimes you need -130 C, and you need LN cans for
longterm storage. Notice that lab freezers rack up ridiculous electricity
bills, and dissipate more heat than an oven, especially in the summer
(SoCal summer + garage + lab freezer = trouble). Of course, you could put
a stainless steel dewar with dry ice into a -15 C freezer, that will give
you a small pocket of ~-80 C, for a bargain price. Especially, if you
pick up the dry ice blocks and crush them by yourself, and don't go for
the long term.

> > c) Sterilizer: $1000-$15000
> alcohol won't do?

No. Cold sterilization solutions are available, but what you need is a
small autoclave for the eppendorf caps and tips (don't forget the
sterilization indicator tape), and for solution flasks too, of course. you
can use a pressure cooker with a little water in it, and a raised platform
in it. Don't bother. Just buy a used desktop autoclave. They're quite
cheap.

> > d) Incubator: $2000-$5000
> 2 grand for a heater!? are you serious? any disadvantage in using the
> "use a lamp" method i read somewhere on the net?

Incubators do not only temperature, but also humidity and gases. You can
improvise here, of course.

> > e) Centrifuge(s): $1000-$7000
> yes, i assume for isolating ribosomes, you'd want tens of thousands of
> RPMs, but just to separate DNA, what would be the minimum RPM
> requirement? wouldn't a souped up fan spun for a long time do? (only
> half kidding)

If you're talking separation in CsCl gradient, that's an ultracentrifuge.
Uh, you don't want to go there. Something requiring a microbalance to
equilibrate the tubes, solid titanium rotors, oildiff evacuated *armored*
rotor chamber is not something you'd want to operate in your home, even if
you can get one for cheap.

> > f) PCR apparatus: $2000-$4000
> on the cheap side, i was thinking of using manpower: 95C tub, 60C tub,
> 72C tub, move eppendorfs from one to the other when you think it's
> appropiate. If this is too mindnumbing and feeling enterprising, grab
> lego mindstorms, get it to do it for you.

Just drill a number of holes into an aluminum block, put it on a Peltier
and drive it by software.

> > f) Microscope: $400+
> i suppose

This one is cheap, but for serious work you'll need EM. Getting a used one
for cheap is easy, but you won't be able to maintain it by yourself.

> > g) Hood: $3000+
> again, is it really necessary? (as opposed to, say, fan pointing out the
> window)

You have a problem there. You need to first learn what the gadgets are
there for, to realize which of the functionality can be sacrificed and
which can be replicated on the cheap. Luckily, it is very possible to
build a laminar flow bench DIY.

> I know it would be a very dodgy setup, but would it be good enough to
> isolate prokaryotic dna, cut it up and gel it? what about eukaryotic?

DNA preps and digests are easy, assuming you have your enzymes (not very
cheap), and work cleanly (and autoclave all of your gear). Agarose gels
can be made in a microwave, and poured into a form.

You can do a lot if you know what you're doing, and shop in used equipment
shops (UCSD has one, and there are others, but prepare for long and
relatively frequent drives when fishing for eqipment).

To learn the basics it would seem to be best to spend some time as an
indentured slave in a molbio lab, doing grunt work. Notice that if you set
up a lab at home, you will be under official scrutiny of the feds,
including little unannounced visits (some of them with a search warrant).

Assuming any of you are crazy enough to want to set up something like this
at home, I'm very ready to offer advice and ideas.



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