J

jot

pipette puller? what kind of ghetto device is that? i was just thinking today - what would i do with out those crazy guys at Quigen? is there any molecular bio done without kits - or has industry thoroughly spoiled me? i was just thinking maybe i should find out exactly what is in "buffer P1, P2, P3", and what exactly the robotic arm in the corner is doing all the time - this place is so rich its making me dumb.
 

wgu

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In my previous academic lab, I lived on a supply line of Minipreps kits. Odd that I'm on the other side of the story right now. I don't work for Qiagen but my company makes immunoassay kits. Buffers are always hush-hush if they have a non-patented ingredient. We even had a rival company look through our trash. Ha ha :)
 
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Doctor&Geek

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Haha,

I think the folks in my lab just said there was some EDTA [metal ion chelator] in those solutions, or something like that. Solution III smells just like acetic acid -- I doubt they even made any changes to it!

Heck, Qiagen is so expensive anyways; just make your own buffers from maniatis. =]

Jason
 

Original

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Jason,
'Sup? I had dinner with ur MSTP classmates but you were MIA. One of them is my homie so I gate-crashed into the dinner with her. They're a nice group. I wish I could have gone to the MSTP retreat with them.
 

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I'd definitely say TEMED is way worse than BME or DTT.

And by the way, I make my own solutions for minipreps; no kit or mini-column materials whatsoever. :) I can pm the recipe/protocol if anyone wants, it's faster than the promega or qiagen one.

I make almost everything from the basic ingredients, although i'm starting to use the NuPage pre-cast gels for westerns and absolutely love'em. Highly recommended, perfect beautiful transfers, low-cost, keep forever.
 

Doctor&Geek

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If you're using Maniatis, I've always found that phenol extraction takes twice as long to finish with more trouble. Usually the product is less clean if you're wanting to do sequencing. That white precipate that shows up sucks and will make your sequencing results phooey. That's why filters are good =].

Of course, if your lab has funds up the wazhoo, you buy your
own water instead of picofiltering and autoclaving it =], let alone getting precast gels.

Jason
 
J

jot

hah - at supa rich industry everything is one-use/disposable - from the water as someone mentioned to the glass flasks (they just remelt and redeliver) -- even people were even slightly cost concious there millions would be saved. before they had their little stock snafoo i had a $1000/day spending limit without authorization needed (cause they can write it off in taxes, anything greater than that is considered capital expense). i've never seen scientists mood so dependent on a stock price. score- time to head to work.
 

Neuronix

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i was just thinking maybe i should find out exactly what is in "buffer P1, P2, P3", and what exactly the robotic arm in the corner is doing all the time - this place is so rich its making me dumb.

...

I think the folks in my lab just said there was some EDTA [metal ion chelator] in those solutions, or something like that. Solution III smells just like acetic acid -- I doubt they even made any changes to it!

You guys must be talking about extracting your minipreps of plasmid DNA from bacterial colonies. Here you go:

Solution 1:
50mM glucose
25mM Tris.Cl (pH ~8.0)
10mM EDTA

Solution 2:
.2N NaOH
1% SDS
H20 (to reduce NaOH concentration from your stock)

Solution 3:
3M Na Acetate OR 5M K Acetate (pH ~5.0)
Glacial acetic acid
H20

(This is from our lab manual, I'm at home so I don't have the proper name to credit it. If you really needed it I could figure out how many mL for each, but my figures are back in lab too)

The little robotic arm might be part of a PCR machine, some people are especially finicky about their PCRs and don't want the temperature ramping up and down while cycling and all that. Maybe not tho, who knows.
 

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Originally posted by jot
pipette puller? what kind of ghetto device is that? i was just thinking today - what would i do with out those crazy guys at Quigen? is there any molecular bio done without kits - or has industry thoroughly spoiled me? i was just thinking maybe i should find out exactly what is in "buffer P1, P2, P3", and what exactly the robotic arm in the corner is doing all the time - this place is so rich its making me dumb.

Yes, our generation of laboratory labor is HIGHLY spoiled....a postdoc in my group likes to rant on about the days they used to do PCR reactions BEFORE THE THERMOCYCLER.

Could you imagine sitting in a room for three hours, moving tubes between three different waterbaths going "heat, anneal, polymerize, heat, anneal, polymerize" 30 times over? Sounds so boring it makes my skin crawl.
 
J

jot

nah - the robotics arm does immunostaining i guess - which is funny because it breaks about 20% of the slides which take an incredible amount of effort to prepare (surgery/sectioning/fixing). these pcr machines get smaller and smaller (the new one here is the size of a 3.5 disk rack holder thing)- i dunno how they can ramp up and down so fast - thats something for the heat transfer boys ...
 

exigente chica

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I aslo am in a summer lab that doesn't beleive in kits, so I make my own solutions too bikini pricess:p
 

Doctor&Geek

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***********
these pcr machines get smaller and smaller (the new one here is the size of a 3.5 disk rack holder thing)- i dunno how they can ramp up and down so fast - thats something for the heat transfer boys ...
***********

I believe that in physics there is something called the Peltier effect -- using two different metals of different thermoconducting capabilities to quickly heat or cool one of those metals. Don't ask me about it -- all I know is that it's used in CPU heatsinks =]

Jason
 
J

jot

yeah - about those chips - we just got a new machine that does pcr on a chip in order to capitalize on those sharper heat transfer gradients -- now if only we could think of a good reason we bought it ....
 
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