Bad day in the Lab rant

This forum made possible through the generous support of SDN members, donors, and sponsors. Thank you.
BlueElmo

BlueElmo

Full Member
15+ Year Member
Joined
Sep 7, 2006
Messages
14,411
Reaction score
26
So we were doing RNA isolation in my research lab today, and I made two really really stupid mistakes. The protocol called for me to add 100 uL of chloroform to each tube of RNA but I went and added 1 mL.
It also called for me to centrifuge at 12,000 g for 15 minutes and I centrifuged it at 20,000 g. The result was complete disaster: could get no RNA isolation and two tubes completey broke in the centrifue machine, losing all the samples.
I think I was totally out of it when I was doing this. I'm an idiot.

The result was that bunch of people, including my advisor, in the lab had to spend couple hours trying to fix my mistakes. It would have been funny if I hadn't seen the pure murder in their eyes...😳
There goes my hope for a good LOR from my advisor. Sigh.
Members don't see this ad.
 
i see why you're blue
 
I suspect that your advisor has watched many, many students screw in up lab and isn't really going to hold it against you all that much. Lab mistakes happen. And in the grand scheme of things, yours is actually very minor 😛
 
Members don't see this ad :)
BlueElmo said:
So we were doing RNA isolation in my research lab today, and I made two really really stupid mistakes. The protocol called for me to add 100 uL of chloroform to each tube of RNA but I went and added 1 mL.
It also called for me to centrifuge at 12,000 g for 15 minutes and I centrifuged it at 20,000 g. The result was complete disaster: could get no RNA isolation and two tubes completey broke in the centrifue machine, losing all the samples.
I think I was totally out of it when I was doing this. I'm an idiot.

The result was that bunch of people, including my advisor, in the lab had to spend couple hours trying to fix my mistakes. It would have been funny if I hadn't seen the pure murder in their eyes...😳
There goes my hope for a good LOR from my advisor. Sigh.
Click to expand...

You'll be fine, provided you cleaned the centrifuge, then cleaned everyone's cars and had their shoes polished, and then picked up their dry cleaning...

Lol... I think at some point, every single one of us does something truly stupid in the lab. My advise? Drink more coffee, rock out a lot of excellent samples.
 
I just had a whole month of my work lost so I understand how you feel. It sucks but I am more determined to do it right this time.
 
hey i had same thing too... my very first chem lab was on friday and i broke a 800 mL beaker... like 5 mins after my partner broke a thermometer 🙁
 
That brings to mind this organic lab I took. We were extracting caffeine using some volatile solvent (hexanes, I think). We put the solvent with some heated solution containing caffeine, sealed the mixture in a tube, and centrifuged. Half the classes' tubes exploded in the centrifuge... and the other half while they were trying to decant the supernatant. XD!

But like the others said, I really wouldn't worry. Mistakes happen. Laugh it off and improve next time. 🙂
 
Thanks.
I feel really bad though because apparently they were preparing those samples this entire winter break (I was off on my own vacation and didn't show up).
And here I come blow it all off in 20 minutes. LOL...🙁
 
Being that this is my first post, i'll get it out of the way and introduce my stupidity along with myself.

This was like my first or second protein purification and i had spent all day growing, then inducing my bacteria. So finally i am ready to spin down and go home, but of course in my hurry to get out of there i forgot to switch the cultures from the glass flasks to centrifuge tubes. Around 4000xg they all exploded (except one - causing the centrifuge to almost jump off the desk). i was pretty sure that i had ruined it (though if i had, i probably would have been the hero of the floor. This thing was a pos - my PI bought me a new one not long after...got to love the start-up fund😀).

Luckily, this was late at night and no one was around to witness such idiocy.

Anyways, don't worry about it. I can guarantee your PI has screwed up just about everything possible in your lab. Just learn your lesson, and don't let it happen again. It's amazing how impressive you will seem if you learn your lesson the first time. After a couple of months you'll be a stud.

Then again, if you are a couple of months in and this is kind of thing is your MO, you might want to start a habit of double checking your protocols before you act.
 
Last edited:
i suck at posting too
 
Last edited:
I dropped a really fat column on the floor, and it was filled with all kinds of goodies for me to clean up. ugh
 
Members don't see this ad :)
If you do a search you'll find an entire thread dedicated to the various screw ups people have done in lab.

As long as you didn't break the ultracentrifuge or blow up the wing of your building you'll be fine. As long as you learn from your mistake and don't repeat it again.

In my school someone dropped an NMR tube into the NMR without the little holder/mirror thing that floats in the current of air. People get VERY angry when you have to purge the magnet so you can go in there and clean it out.
 
Last edited:
BlueElmo said:
There goes my hope for a good LOR from my advisor. Sigh.
Click to expand...

I hope you aren't just doing this research for a recc. If that's the case you will keep making mistakes simply because you don't give a damn about your research.
 
cyclin M said:
I hope you aren't just doing this research for a recc. If that's the case you will keep making mistakes simply because you don't give a damn about your research.
Click to expand...

I'm not. It was just an afterthought.
 
BlueElmo said:
I'm not. It was just an afterthought.
Click to expand...

yeah I was just kidding elmo. You're gonna be fine. Everyone makes mistakes, some big some small. An example of a big one is a huge fire that chars a quarter of the lab and half the lab's experiments with it, and a small one is losing an RNA sample which will take you probably a week at most to get again with another culture and whatnot. Don't let this mistake define you and don't let it kill your interest in research. 👍
 
personally i say its the PI's fault for entrusting a major project like this in the hands of an undergrad, but even worse, a pre-med undergrad to boot
 
Lukkie said:
personally i say its the PI's fault for entrusting a major project like this in the hands of an undergrad, but even worse, a pre-med undergrad to boot
Click to expand...

Why is it wrong to entrust projects to pre-med undergrads? His PI clearly saw something in him.
 
Lukkie said:
personally i say its the PI's fault for entrusting a major project like this in the hands of an undergrad, but even worse, a pre-med undergrad to boot
Click to expand...

Not at all. There are plenty of undergrads that get their own projects. Many have publications to show for it. I have had a lot of independence in my lab activities and I've been ever thankful for it.
 
xanthomondo said:
If you do a search you'll find an entire thread dedicated to the various screw ups people have done in lab.

As long as you didn't break the ultracentrifuge or blow up the wing of your building you'll be fine. As long as you don't repeat your mistake and repeat it again.
Click to expand...

Yeah I hear departments get a little angry when the huge ultracentrifuges end up breaking and fly across rooms.
 
Dude that's nothing... you messed up one experiment. Yeah, it took a lot of time to prepare it, but that's basic research for you. This kind of stuff happens, and as long as you learn from your mistake your advisor really won't care. People have done worse before you, and they'll do worse after you. It'll be funny in about 3 weeks.
 
I have a friend who worked in a lab for about a year in college. He worked on a big study that ended up a bust.

In med school mol bio lab, I noticed that he was using the pipette wrong and basically putting massive quantities of reagents in everything.

A look of horror flashed across his face when I showed him how you're supposed to use the thing...:laugh:
 
Personally, I think that RNA isolation tempts accidents and I have seen MANY of them during my lab time.

I have been personally responsible for losing RNA from a rare-cell line (I was separating the cells, and I sneezed). However, the worst I have seen was an undergrad in training - this girl was extremely nervous and actually managed to spill the bottle of chloroform on the PIs arm when he came over to check on her. Yeah, very poor form.

At least your mistake could be fixed in one afternoon. Trust me, everyone will forget all about a slip up like that with time.
 
stb said:
Being that this is my first post, i'll get it out of the way and introduce my stupidity along with myself.

This was like my first or second protein purification and i had spent all day growing, then inducing my bacteria. So finally i am ready to spin down and go home, but of course in my hurry to get out of there i forgot to switch the cultures from the glass flasks to centrifuge tubes. Around 4000xg they all exploded (except one - causing the centrifuge to almost jump off the desk). i was pretty sure that i had ruined it (though if i had, i probably would have been the hero of the floor. This thing was a pos - my PI bought me a new one not long after...got to love the start-up fund😀).

Luckily, this was late at night and no one was around to witness such idiocy.

Anyways, don't worry about it. I can guarantee your PI has screwed up just about everything possible in your lab. Just learn your lesson, and don't let it happen again. It's amazing how impressive you will seem if you learn your lesson the first time. After a couple of months you'll be a stud.

Then again, if you are a couple of months in and this is kind of thing is your MO, you might want to start a habit of double checking your protocols before you act.
Click to expand...

luckily the actual centrifuge didn't break, then you'd be screwed
(thats one of my nightmares.. me breaking the centrifuge that is worth thousands)

i do protein purification too.. isn't it a big pain? especially the part where you try to put the protein pellet back into solution.. my arm physically starts to hurt pipetting up and down and up and down. :/
 
I don't make mistakes in my lab.
 
BlueElmo said:
So we were doing RNA isolation in my research lab today, and I made two really really stupid mistakes. The protocol called for me to add 100 uL of chloroform to each tube of RNA but I went and added 1 mL.
It also called for me to centrifuge at 12,000 g for 15 minutes and I centrifuged it at 20,000 g. The result was complete disaster: could get no RNA isolation and two tubes completey broke in the centrifue machine, losing all the samples.
I think I was totally out of it when I was doing this. I'm an idiot.

The result was that bunch of people, including my advisor, in the lab had to spend couple hours trying to fix my mistakes. It would have been funny if I hadn't seen the pure murder in their eyes...😳
There goes my hope for a good LOR from my advisor. Sigh.
Click to expand...

If it's any consolation, I once forgot to completely tighten the door on an industrial sized autoclave and essentially turned the lab into a sauna while as at lunch. It ruined dozens of lab books and materials all over the lab. I was mercilessly made fun of for months, but eventually got back into the good graces of my PI.
 
Last edited:
How many samples were you running? I'm more than familiar with rna isolation and we always tend to run experiments in duplicate/triplicate so that when we isolate the rna, we get the yields we want and don't have to worry about running the experiment again if that happens.


I had a similiar issue, tho, so don't sweat it. I had the samples in trizol and added the chloroform but forgot to shake the samples up and just centrifuged it:laugh:
 
lord_jeebus said:
I have a friend who worked in a lab for about a year in college. He worked on a big study that ended up a bust.

In med school mol bio lab, I noticed that he was using the pipette wrong and basically putting massive quantities of reagents in everything.

A look of horror flashed across his face when I showed him how you're supposed to use the thing...:laugh:
Click to expand...

😱
 
URHere said:
Personally, I think that RNA isolation tempts accidents and I have seen MANY of them during my lab time.

I have been personally responsible for losing RNA from a rare-cell line (I was separating the cells, and I sneezed). However, the worst I have seen was an undergrad in training - this girl was extremely nervous and actually managed to spill the bottle of chloroform on the PIs arm when he came over to check on her. Yeah, very poor form.

At least your mistake could be fixed in one afternoon. Trust me, everyone will forget all about a slip up like that with time.
Click to expand...
I'm just wondering why people still use chloroform/ethanol/water extraction methods when there are kits made by companies (qiagen) that yield much purer rna without the foul/dangerous reagants.

I'm only asking this because I'm curious if it's a cost issue or simply a lack of trust in newer methods
 
One time I was putting bacteria onto an agar plate (I forgot what that procedure is called - transformation?). To do that, you have to sterilize the thing you use to plate with (I'll call it a "plater"), usually with an alcohol mixture. You dip the plater in the alcohol mix (i.e. part water, part ethanol), and then before put it on the plate, you put the tip against a bunsen burner. Well, we ran out of the alcohol mix, so I made some 50% solution I believe.

After I did the plating, I put the plater back in the alcohol mix and it caught on fire. Fortunately, another lab member was there who brilliantly suggested to cover the beaker with foil until the fire died. Then, I stupidly tried to dump the alcohol mix out, but discovered it was rather hot and dropped the beaker.

So on the first day of unsupervised work, I almost set the lab on fire and broke a beaker.

Good times...
 
Kaustikos said:
I'm just wondering why people still use chloroform/ethanol/water extraction methods when there are kits made by companies (qiagen) that yield much purer rna without the foul/dangerous reagants.

I'm only asking this because I'm curious if it's a cost issue or simply a lack of trust in newer methods
Click to expand...

Qiagen kits are the bomb.
 
xanthomondo said:
If you do a search you'll find an entire thread dedicated to the various screw ups people have done in lab.

As long as you didn't break the ultracentrifuge or blow up the wing of your building you'll be fine. As long as you learn from your mistake and don't repeat it again.

In my school someone dropped an NMR tube into the NMR without the little holder/mirror thing that floats in the current of air. People get VERY angry when you have to purge the magnet so you can go in there and clean it out.
Click to expand...
I was so scared when using the ultracentrifuge. It took a lot of work to prepare the samples that we needed, and my postdoc's description of the machine seemed like a small mistake could ruin the machine.
 
savant said:
One time I was putting bacteria onto an agar plate (I forgot what that procedure is called - transformation?). To do that, you have to sterilize the thing you use to plate with (I'll call it a "plater"), usually with an alcohol mixture. You dip the plater in the alcohol mix (i.e. part water, part ethanol), and then before put it on the plate, you put the tip against a bunsen burner. Well, we ran out of the alcohol mix, so I made some 50% solution I believe.

After I did the plating, I put the plater back in the alcohol mix and it caught on fire. Fortunately, another lab member was there who brilliantly suggested to cover the beaker with foil until the fire died. Then, I stupidly tried to dump the alcohol mix out, but discovered it was rather hot and dropped the beaker.

So on the first day of unsupervised work, I almost set the lab on fire and broke a beaker.

Good times...
Click to expand...

Sodium hydride fires are always fun to deal with.
 
savant said:
I was so scared when using the ultracentrifuge. It took a lot of work to prepare the samples that we needed, and my postdoc's description of the machine seemed like a small mistake could ruin the machine.
Click to expand...
They do that so you're not careless, imo. I've become so careless sometimes when dealing with certain machines that I sometimes need a reminder that these things cost upwards of 200,000 each. But then they talk about putting these things in salvage for 10 years because no one's going to use them and just upgrading the units like it's nothing. Luckily no machines have broken under my use. *knocks on wood*

I just know that transitioning from a pharmaceutical company to graduate school would be brutal....:laugh:
 
I lost a MaxiPrep sample today....dumped the eluate into the wash buffer flow-through. Then tried to salvage the DNA by doing multiple Isopropanol/70% EtOH extraction...I shall see tomorrow if it worked :laugh:
 
chessodoc said:
I lost a MaxiPrep sample today....dumped the eluate into the wash buffer flow-through. Then tried to salvage the DNA by doing multiple Isopropanol/70% EtOH extraction...I shall see tomorrow if it worked :laugh:
Click to expand...
I always liked doing MaxiPreps for some reason. Maybe the excitement of having a lethal dose of EtBr
 
Kaustikos said:
How many samples were you running? I'm more than familiar with rna isolation and we always tend to run experiments in duplicate/triplicate so that when we isolate the rna, we get the yields we want and don't have to worry about running the experiment again if that happens.


I had a similiar issue, tho, so don't sweat it. I had the samples in trizol and added the chloroform but forgot to shake the samples up and just centrifuged it:laugh:
Click to expand...

I was running 22 samples. But like I said, the samples looked so weird when I added 1 mL of chloroform. I mean, this biconcave shaped/red-blood cell shaped thingies were floating on top. :laugh:
 
gatorsbball said:
Really? Maxipreps were always a chore for me.. too much waiting around 😴
Click to expand...
Eh.. minipreps were too boring.

At least with maxipreps you get to crimp the tubes and use syringes to draw out the suspended section of fluid.

All the maxipreps I did, we left the ultracentrifugation overnight, so I don't know if that's the wait you were referring to...
 
Brodiewankenobi said:
no biggie if you were doing an ethanol extraction. The chloroform should just evaporate. However, if you were doing a TRIzol extraction, that could actually be kinda dangerous.

http://en.wikipedia.org/wiki/Trizol
Click to expand...
Pssht. I use trizol in almost all my extractions and even spray a little in the morning as my cologne. I like to use it in none-fume hoods, that way everyone gets the sedating effect.
 
gatorsbball said:
Really? Maxipreps were always a chore for me.. too much waiting around 😴
Click to expand...

savant said:
Eh.. minipreps were too boring.

At least with maxipreps you get to crimp the tubes and use syringes to draw out the suspended section of fluid.

All the maxipreps I did, we left the ultracentrifugation overnight, so I don't know if that's the wait you were referring to...
Click to expand...
This is what I use now: put samples in and come back 45 minutes later.

200732893740.jpg
 
This is what I use now: put burritos in and come back 1 minute later.

sanyo_microwave.jpg
 
norgen kits >> qiagen 🙂
 
yeah me too
 
You must log in or register to reply here.

Similar threads

PickaGodnPray
Rant
Replies
17
Views
3K
PickaGodnPray
PickaGodnPray
C
Is it a bad look to not attend a second look day?
Replies
8
Views
1K
Mr.Smile12
Mr.Smile12
R
  • Question Question
is it rude to ask shadowing the day of?
Replies
5
Views
917
GoSpursGo
GoSpursGo
I
can i use the phrase until the day I die in my PS
Replies
8
Views
1K
infinitestars
I
kv11
  • Poll Poll
Wet lab vs Clinical Research
Replies
5
Views
1K
pinkdino
P
Top