Poll-What is your least favorite lab technique?

[Treg]

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After about 1000 PCRs to screen transgenic mice, I think I would rather have an EtBr slurpee than do another.

Also, Southern blots are right up there. Gee, let's spend a week working on something, only to find out it didn't work.

What about you?

 

[coldchemist]

I have to do a lot of image processing via computer. Personally, I'd much rather be purifying proteins.

 

[UDbiochem]

replacing the membrane on an oxygen electrode.... what a pain in the you know what

1. remove old membrane
2. scrape algae off electrode surface with small razor blade
3. place drop of (potassium chloride?) solution on tip of electrode
4. remove membrane from package, carefully place the membrane over electrode tip, make sure to capture solution drop between membrane and platinum electrode tip.
5. if you lose the drop, you have to repeat the whole process AGAIN

Ditto about the southern blots too, extremely annoying as well

 

[superdevil]

although it sounds harmless enough, the process of pressure-filtering 15 liters of media for cell cultures is a long, boring, labor-intensive way to waste an hour and a half. i wish my lab had the cash to just BUY the friggin' stuff instead

😡 😡

also, any time where you are trying to find the proper optimization of pretty much any particular protocol (like running a dozen transient plasmid transfections because your cells are wussy, picky little sluts!) will cause frustration.

 

[pathdr2b]

The cervical dislocation of large rodents.

 

[Treg]

I refuse to do cervical dislocation ever since the following incident (warning-not for squeamish)

Autoimmune diabetic mice have vasular problems, resulting in weakened skin. I had a diabetic to sacrifice, and the whole tail ripped off, and the mouse walked away with a bloody gaping hole in its rear end.

The trick is to use dry ice + water, and leave it to RIP in the fume hood.

😱

 

[Pinkertinkle]

Getting the spleen from a mouse you injected proteins into is rather disgusting.

 

[Fritz]

PCR is definetly #1 most annoying technique.

 

[dan0909]

bleh

 

[Primate]

You should try it with subjects from the dog colony. 😱

Seriously, making embryonic chick/mouse extract is perhaps the most tedious, mind-numbing, PAIN IN THE BUTT (next to viral preps). 😉

P

 

[Naphtali]

Ha! I laugh! I trust none of you have ever done Boyden Chamber motillity assays. Coat the bottom chamber with FBS overnight. Load in media of choice. Now lay a membrane over the lower chamber-- and wells with air bubbles are invalid. Now clamp the top chamber on. Fill with cells and media. If it leaks (not uncommon), the whole Boyden is bunk.

PI: "So how's the Boyden going?"
Student: "I didn't pan out. I'm going home to... uh... read some articles. Yeah, that's it."

Also, I once did a Western, and it was PERFECT. And then the secondary antibody didn't work. (Someone left it out on the bench overnight, I later learned.)

Naphtali

 

[dan0909]

I HAVE used Boyden chambers before...yes, they are a bit tedious, but then we switched over to 24-well plates.

 

[Bad Mojo]

Working in a yeast lab really handicaps me in this one. I have not yet taken the kidney or something out of a yeast tetrad. Any technique can really stink if you have to do it over and over and over. With that said, 96-well anything is great (sorry, slightly off topic). It makes me feel super-powerful to control so much data.

 

[Pinkertinkle]

What's with all the PCR haters, all ya gots to do is mix it up and leave it in the machine for 3-5 hours. I had to make some competent bacteria today, and thats a bitch when my bacteria end up incompetent.

 

[surge]

French pressing 2L of bacteria with a 50 mL cylinder.
I'm hoping most of you don't know what the hell I'm talking about.

Ah, undergraduate research... 😀

 

[Primate]

I've always found PCR to be satisfying in some way. It's so Betty Crocker - just mix and go have four or five coffees while it cooks up.

As far as migration assays, I think I have the chamber beat. I developed a prep where we had to dissect out the forebrain (cortex only) of E15 mice (timing the pregnancies - first problem), grow them in various conditions for about a week until we got (or didn't get - the second problem) axons in the dish. Next we had to take the brains of E12.5 and E15 (problem 3) mice and dissect out just the lateral or medial ganglionic eminence (problem 4), dissociate them, label them with flourescent dye (BIG problem), add them to the first culture, let them adhere to axons (hopefully), put them on a scope, identify candidate migrators, image each cell for hours, analyze the data.

If anything went wrong (up to the analyze part) in either set of dissections or timing of pregnancies, the WHOLE prep was kaput and we had to start over. That once happened 5 times in a row. Different problem each time, for over a month out of my life. And that was after the assay was "working" well. I liked the surgeries and such, but the waiting was terrible. Well, that's science. At least we had a nice view out the lab windows. 🙂

 

[R_C_Hutchinson]

the ordering of pcr primers as a process has full and unrestricted permission to lick my left testicle

 

[strangeattractor]

nonlinear unidentifiable model fitting. the problem is there's really no true way of proving a model is unidentifiable if it's nonlinear, and it might even be locally identifiable if you know the rest of the parameters in the right range, given that bovine hormone specific activity molar equivalency ratio you found will work for human hormone (usually it doesn't) and the work of others that hasn't been repeated since 1967 is reliable. let alone initial values, sheesh. and even THEN, what kind of algorithms do the most helpful programs have? some pretty basic wrss equation that barely allows you to modify specific data set weighting. and then, who's to know what kind of convergence criterion you need to aim for at first??

hee hee! obviously i don't work in a wet lab...

 

[Treg]

Today I would rather do anything than pH buffer. Also, regarding buffer, my supervisor has a certain craziness that is as follows:

Wash 9x tap water
Wash 9x DI water
Wash 9x MQ water
All with vigorous shaking in between, of course.

Did I mention that the bottles have been washed and autoclaved by the wash-up facility beforehand?

If the buffer bottle is not washed as such, prior to autoclaving with buffer in it, the quality of the buffer will be highly questionable.

Please God don't let me turn out like this!!😀

 

[vixenell]

Mounting tissue sections.

::shudders::

 

[autobot]

Ovariectemizing 30 mice in one sitting. The reward is definitely seeing all of them wake up.

 

[Airborne]

Better yet -

Bile duct ligating on-told amounts on rats to have them wake up and slowly cirrhose over a period of weeks while you feed and "check-up" on them daily to ensure they have "passed-on" and eventually come to near death before they are sacrificed -

Ah yes, in the name of science.

 

[BandGeek]

Try ovaectimizing four sheep in a day.

I really hate image analysis. I lost 15 pounds in one summer from puking after sitting at a scope for four hours at a time.

Ribonuclease Protections Assays with P32 are pain as well.

 

[KBK]

Originally posted by surge
French pressing 2L of bacteria with a 50 mL cylinder.
I'm hoping most of you don't know what the hell I'm talking about.

Ah, undergraduate research... 😀

Yeah....french pressing definitely sucks. Especially when you spray bacterial juice everywhere....

Also, there is nothing worse than your girlfriend waking you up at 3am so that you can help her dissect 12 mice...

 

[vixenell]

Originally posted by autobot
Ovariectemizing 30 mice in one sitting. The reward is definitely seeing all of them wake up.

Forgot about that one.

 

[Xega]

Originally posted by Treg
I refuse to do cervical dislocation ever since the following incident (warning-not for squeamish)

Autoimmune diabetic mice have vasular problems, resulting in weakened skin. I had a diabetic to sacrifice, and the whole tail ripped off, and the mouse walked away with a bloody gaping hole in its rear end.

The trick is to use dry ice + water, and leave it to RIP in the fume hood.

😱

Ugh I had the same thing happen to me with my first cervical dislocation. I refused to do any more for a while, finally had to one night when no one was around (can't use the dry ice method since I was studying levels of some brain proteins and that is enough time to get changes from asphyxiation).