Bad day in the Lab rant

[Excelsius]

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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...🙁

While accidents do happen, in your case, you were not prepared it seems. It largely depends whether you took your vacation on your own or whether your teammates in the lab wanted you to take some off time based on your hard work in the lab. I am just starting research in a medical school and after talking to the head and learning how hard it is, I know that there is a definite commitment requirement. I was honestly told that without a part time commitment every single week, this is not going to work. If they don't see significant improvement within a month, then you're just not cut out for that type of research. There are also several books I am reading on my own before even the research starts. The point here is if you don't care for research, you shouldn't do it. Researchers work very hard to get grant money for the research. One of my teachers hadn't slept all weekend writing a grant proposal. A little more care and respect can go a long way. This is not just about your letter of recommendation.

Ask yourself why you made not one but two mistakes within a few minutes. If you don't like the research, don't do it. Actually you might find a research project that interests you more, but if you don't, that's ok. Just make up your time somewhere else - leadership, volunteering, etc. Sorry if I sound a bit dry, but I have been working in a place where research can save or endanger lives. Seeing some guys who already have their BS and are doing research without any deep understanding as to what they are doing has been a disappointing experience - they really should not have made it through. Premeds have such a bad name in many facilities because professors know that most of the herd is in it for quantity, not quality. As a result, anyone who is a traditional premed already has a negative sign to next to his name. What you guys do affect all of us. Good luck.

 

[vadd0]

While accidents do happen, in your case, you were not prepared it seems. It largely depends whether you took your vacation on your own or whether your teammates in the lab wanted you to take some off time based on your hard work in the lab. I am just starting research in a medical school and after talking to the head and learning how hard it is, I know that there is a definite commitment requirement. I was honestly told that without a part time commitment every single week, this is not going to work. If they don't see significant improvement within a month, then you're just not cut out for that type of research. There are also several books I am reading on my own before even the research starts. The point here is if you don't care for research, you shouldn't do it. Researchers work very hard to get grant money for the research. One of my teachers hadn't slept all weekend writing a grant proposal. A little more care and respect can go a long way. This is not just about your letter of recommendation.

Ask yourself why you made not one but two mistakes within a few minutes. If you don't like the research, don't do it. Actually you might find a research project that interests you more, but if you don't, that's ok. Just make up your time somewhere else - leadership, volunteering, etc. Sorry if I sound a bit dry, but I have been working in a place where research can save or endanger lives. Seeing some guys who already have their BS and are doing research without any deep understanding as to what they are doing has been a disappointing experience - they really should not have made it through. Premeds have such a bad name in many facilities because professors know that most of the herd is in it for quantity, not quality. As a result, anyone who is a traditional premed already has a negative sign to next to his name. What you guys do affect all of us. Good luck.

tl;dr.

 

[cyclin M]

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.

please show me this magical bottle that can "spray" viscous fluid.

 

[notaboutthebike]

...Maybe the excitement of having a lethal dose of EtBr

If you didn't lol at this, you either haven't had enough lab experience yet or you haven't had a neurotic lab supervisor that acted like it was a mixture of anthrax and kryptonite.

 

[TMP-SMX]

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.

Wha? I always used the trizol with no fume hood. I actually got a little spray on my face from the micropipette once. It burned a bunch of little tiny holes in my cheek.

 

[Lukkie]

tl;dr.

it was a good post vadd0 😳

 

[BlueElmo]

tl;dr.

what does that mean?

 

[paki20]

dude thats nothing...you'll be fine...just apologize again to them tomorrow..my f*ck up is a lot worse:

it was my first time collecting a kidney biopsy alone (the doctor obviously performed the biopsy..i was just there to collect a small piece)..well i thought the doctor cut the piece for me so i ended up taking the entire kidney biopsy and placed it in liquid nitrogen to analyze for research purposes..leaving nothing behind for the pathologists to analyze in order to make a diagnosis for the patient...and this biopsy was CRUCIAL because the patient was potentially experiencing rejection..the doctor was SOO pissed..i got yelled in front of all the nurses...i just profusely apologized many times..my PI from lab was pretty upset also..had an emergency meeting with the ENTIRE lab staff at 7 am the following day...luckily the biopsy was still viable for the pathologists to analyze..phew

but yeah there goes my LOR..haha

 

[Lukkie]

what does that mean?

secret code only johns hopkins acceptees can decipher

 

[Marjan Islam]

if this makes your feel any better, I forgot to turn in some important forms which withheld my ENTIRE lab from getting new animal shipments for a week! I still plan on asking my PI for a rec 😛

 

[GoSpursGo]

what does that mean?

Or it was Vadd banging his head on the keyboard...

Seriously, I wonder why some people feel the need to get up on a high horse to talk down at other people. Is it just to make themselves feel better and assure themselves, "Well *I* certainly would never have made such a foolish mistake because *I* am constantly prepared."

I've got a bunch of friends going on to get their PhD's in chemistry, bio, physics, etc... ALL of them have made stupid mistakes in lab. It's not a matter of a premed not caring about his/her research, it's a matter that any undergrad student is fundamentally unprepared to do research at a high level; thus, a PI should not be surprised when an undergrad of any major or future professional plan screws something up once in a while- it comes with the territory of trying to do research at an undergrad institution. This one mistake does not, and should not, define the OP. Furthermore, you can't blame an undergrad student for going home over the holidays- the PI and grad students are getting paid to do research, the undergrad is not; to ask that sort of a commitment at that stage is a little unrealistic. Part of undergrad is actually getting out and LIVING, not spending every waking moment worrying about what will help your application down the road.

**** happens. It's what happens next that defines you.

 

[Kaustikos]

please show me this magical bottle that can "spray" viscous fluid.

Wha? I always used the trizol with no fume hood. I actually got a little spray on my face from the micropipette once. It burned a bunch of little tiny holes in my cheek.

WOW

That joke went right over BOTH your heads.

Of course I use Trizol in a fumehood.

 

[Kaustikos]

dude thats nothing...you'll be fine...just apologize again to them tomorrow..my f*ck up is a lot worse:

it was my first time collecting a kidney biopsy alone (the doctor obviously performed the biopsy..i was just there to collect a small piece)..well i thought the doctor cut the piece for me so i ended up taking the entire kidney biopsy and placed it in liquid nitrogen to analyze for research purposes..leaving nothing behind for the pathologists to analyze in order to make a diagnosis for the patient...and this biopsy was CRUCIAL because the patient was potentially experiencing rejection..the doctor was SOO pissed..i got yelled in front of all the nurses...i just profusely apologized many times..my PI from lab was pretty upset also..had an emergency meeting with the ENTIRE lab staff at 7 am the following day...luckily the biopsy was still viable for the pathologists to analyze..phew

but yeah there goes my LOR..haha

I would've done that too..... At least you put it in liquid nitrogen...I mean, some people I work with think that putting samples in the freezer is fine for long-term storage.🙄

That sucks though

 

[Brodiewankenobi]

WOW

That joke went right over BOTH your heads.

Of course I use Trizol in a fumehood.

hahaha...sometimes the SDN crowd is too serious for me. Given trizols remarkable ability to bind and stablize nucleic acids during homogenization i imagine that it could be quite a mutagen.

 

[TMP-SMX]

WOW

That joke went right over BOTH your heads.

Of course I use Trizol in a fumehood.

It was obvious you were using sarcasm, but I was confused as whenever I was adding trizol I don't recall ever using a fume hood during addition of the trizol. Notice how I said that I always used trizol with no fume hood when you were being sarcastic about your magical spray bottle and sedating the room by not using trizol in a fume hood. I do remember using the hood for other parts of the extraction though so that is probably where the danger of evaporation is. Though this was 2 years ago so I probably forgot or it my brain cells have been dissolved by trizol.

 

[parafilm]

Speaking of bad days in lab... I was almost done with a Maxiprep and then threw out my eluted DNA. Brilliant. My grad student is currently pretending he doesn't want to kill me.

/end rant/

 

[ngkats]

While accidents do happen, in your case, you were not prepared it seems. It largely depends whether you took your vacation on your own or whether your teammates in the lab wanted you to take some off time based on your hard work in the lab. I am just starting research in a medical school and after talking to the head and learning how hard it is, I know that there is a definite commitment requirement. I was honestly told that without a part time commitment every single week, this is not going to work. If they don't see significant improvement within a month, then you're just not cut out for that type of research. There are also several books I am reading on my own before even the research starts. The point here is if you don't care for research, you shouldn't do it. Researchers work very hard to get grant money for the research. One of my teachers hadn't slept all weekend writing a grant proposal. A little more care and respect can go a long way. This is not just about your letter of recommendation.

Ask yourself why you made not one but two mistakes within a few minutes. If you don't like the research, don't do it. Actually you might find a research project that interests you more, but if you don't, that's ok. Just make up your time somewhere else - leadership, volunteering, etc. Sorry if I sound a bit dry, but I have been working in a place where research can save or endanger lives. Seeing some guys who already have their BS and are doing research without any deep understanding as to what they are doing has been a disappointing experience - they really should not have made it through. Premeds have such a bad name in many facilities because professors know that most of the herd is in it for quantity, not quality. As a result, anyone who is a traditional premed already has a negative sign to next to his name. What you guys do affect all of us. Good luck.

Don't make mistakes. Be serious.

But this guy's post is a little annoying.

 

[chessodoc]

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...

Thats cesium prep. I HATE cesium preps...there are so many ways you can die...

1) Large amounts of EtBR --> instant cancer --> slow + painful death
2) Imploding/Exploding Ultracentrifuge --> projectiles traveling faster than bullets --> instant death if near it
3) Imploding/Exploding Ultracentrifuge --> projectiles traveling faster than bullets --> instant death when your PI gets "home"
4) Imploding/Exploding Ultracentrifuge -->projectiles traveling faster than bullets --> slow + painful death at the hands of your co-workers.

 

[Kaustikos]

It was obvious you were using sarcasm, but I was confused as whenever I was adding trizol I don't recall ever using a fume hood during addition of the trizol. Notice how I said that I always used trizol with no fume hood when you were being sarcastic about your magical spray bottle and sedating the room by not using trizol in a fume hood. I do remember using the hood for other parts of the extraction though so that is probably where the danger of evaporation is. Though this was 2 years ago so I probably forgot or it my brain cells have been dissolved by trizol.

I think Trizol's affected us all

 

[BlueElmo]

The PI told me to take the semester off, aka kicked me out. Damn, was my mistake that bad? Depressed.🙁

 

[chessodoc]

The PI told me to take the semester off, aka kicked me out. Damn, was my mistake that bad? Depressed.🙁

Oh...oh wow....too rough, I did that so many times.

 

[NoMoreAMCAS]

I hate labs, completely pointless. I learn everything I need to know in class.

 

[Siverhideo1985]

The PI told me to take the semester off, aka kicked me out. Damn, was my mistake that bad? Depressed.🙁

Serious? For breaking a couple tubes in a centrifuge? He needs to relax a little.

I hate labs, completely pointless. I learn everything I need to know in class.

I love labs....especially working in one, for once I get paid to learn.

 

[NoMoreAMCAS]

I love labs....especially working in one, for once I get paid to learn.

I guess that would be sweet, but i was referring to the mandatory labs we have attached to most of our classes.👎

I don't learn a thing, it's just one big waste of time. They should make it optional so kids interested in research can enjoy them while the rest of us can avoid the pain and agony.

 

[Siverhideo1985]

I guess that would be sweet, but i was referring to the mandatory labs we have attached to most of our classes.👎

I don't learn a thing, it's just one big waste of time. They should make it optional so kids interested in research can enjoy them while the rest of us can avoid the pain and agony.

I agree now that you have clarified.

P.S.: My organic chem lab was awesome though...the T.A. would bring a beer wrapped in toilet paper and teach us stuff like how to synthesize TNT...the chemistry of making very concentrated THC...stuff like that...

 

[GoSpursGo]

The PI told me to take the semester off, aka kicked me out. Damn, was my mistake that bad? Depressed.🙁

That's kinda ridic... definitely an overreaction.

Oh well, what's done is done. Find a new lab next semester and start with a fresh slate.

 

[NoMoreAMCAS]

I agree now that you have clarified.

P.S.: My organic chem lab was awesome though...the T.A. would bring a beer wrapped in toilet paper and teach us stuff like how to synthesize TNT...the chemistry of making very concentrated THC...stuff like that...

Lol, hell yeah. I could deal with a lab like that. If we're gonna be making (or at least learning how to) explosives and chemicals that get you high I might have a much different outlook on lab.

 

[badlabrat]

Everyone is saying "at least you didn't break the huge, thousand dollar centrifuge". Well I'm the girl who did. MY partner and I were so concerned with balancing the centrifuge that I forgot to replace the lid and broke the most expensive centrifuge the lab owns.
Thankfully I have an awesome boss who was thankful that no one was hurt and that I promised to never break anything EVER again.

 

[KTownGT]

Happens to everybody man.

I think the 1mL instead of 100mics was not a travesty. The CIA solution inactivates the RNAse that is added beforehand, more should just react quicker, but still hit a max threshold.

I don't believe it would damage the DNA, much.

The centrifuging at 20k with CIA in it was a mistake though, ha!

EDIT: nvm, RNA extraction. Not sure, but shouldn't it just make the aqueous phase larger?

 

[minerva3121]

Happens to everybody man.

I think the 1mL instead of 100mics was not a travesty. The CIA solution inactivates the RNAse that is added beforehand, more should just react quicker, but still hit a max threshold.

I don't believe it would damage the DNA, much.

The centrifuging at 20k with CIA in it was a mistake though, ha!

EDIT: nvm, RNA extraction. That could have unforeseen consequences. hmm

I'm sure the OP would've appreciated your sympathy if you had offered it two years ago

 

[nysw]

.

 

[Minnerbelle]

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

'Cause it's cheap. If you use Trizol, it can be as cheap as like $1-2/prep. RNeasy kits would run you about $4-5/prep. If you're not doing anything all that fancy with that RNA, a crude extraction method is more than enough. And if you do a LOT of it, it can even save you some time (unwrapping all those columns is pretty cumbersome sometimes. lol).

 

[Truzzi]

Reestablish a positive relationship with your advisor by preparing a mature apology (both written and verbal). Say that you will be certain to be more responsible in the future and that you will make sure to notice the fine details. Beyond that, there is not much you can do.

Aside of baking cookies.

Or bringing her chocolates if she's a female.

 

[cdm3413]

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.

I'm horrible in labs, as well. It's something to do with the hands on + conceptual themes that destroy me. A/A+ in gen. chem lecture 1 and 2, respectively, C+s in all chem. labs.

:\ not a big deal, I'd say, though.

 

[Kaustikos]

Reestablish a positive relationship with your advisor by preparing a mature apology (both written and verbal). Say that you will be certain to be more responsible in the future and that you will make sure to notice the fine details. Beyond that, there is not much you can do.

Aside of baking cookies.

Or bringing her chocolates if she's a female.

Absolutely do this 2 years later! That way the PI will be all "wtf" when they get the chocolates and apology.

 

[kami333]

'Cause it's cheap. If you use Trizol, it can be as cheap as like $1-2/prep. RNeasy kits would run you about $4-5/prep. If you're not doing anything all that fancy with that RNA, a crude extraction method is more than enough. And if you do a LOT of it, it can even save you some time (unwrapping all those columns is pretty cumbersome sometimes. lol).

If you only have a limited amount of sample a Trizol extraction can give you a higher yield as well. If you really want to get the best sample prep, some people swear by doing a Trizol extraction then cleaning it afterwards with the RNeasy cleanup protocol.

My current lab has a Qiacube (automates most of Qiagen's kits, love it for mini preps) but we still do most of our RNA extractions by Trizol.

 

[DNAdude120]

Happens to everybody man.

EDIT: nvm, RNA extraction. Not sure, but shouldn't it just make the aqueous phase larger?

That's what I thought. I figure just running the additional aqueous phase through the column would isolate the RNA well enough.

 

[TheYeren]

I accidentally spilled ethidium bromide all over the supplies in this kid's drawer who was always following me around making sure I didn't screw anything up (he was in the lab for forever and kind of my mentor). Hopefully it doesn't cause any unfortunate mutations for him

Later that day I killed off an entire drosophila line that we had just had specially engineered 🙁

 

[Kaustikos]

That's what I thought. I figure just running the additional aqueous phase through the column would isolate the RNA well enough.

Nope. There is such a thing as TOO MUCH of a good thing. Especially when you add 10x the volume to an extraction protocol.

 

[Kaustikos]

I accidentally spilled ethidium bromide all over the supplies in this kid's drawer who was always following me around making sure I didn't screw anything up (he was in the lab for forever and kind of my mentor). Hopefully it doesn't cause any unfortunate mutations for him

Dude. You're a prick. You're supposed to report **** like that 😡 That's not even close to funny.

 

[TheYeren]

Dude. You're a prick. You're supposed to report **** like that 😡 That's not even close to funny.

Uh I told him about it and we threw everything out... calm down.

 

[KTownGT]

Nope. There is such a thing as TOO MUCH of a good thing. Especially when you add 10x the volume to an extraction protocol.

Hmm, what does the excess Chloroform do to the RNA/DNA? Just curious.

Is it similar to over-drying DNA at the end of a protocol?

 

[Kaustikos]

Uh I told him about it and we threw everything out... calm down.

Uh...you should've clarified that! 😛

 

[Kaustikos]

Hmm, what does the excess Chloroform do to the RNA/DNA? Just curious.

Is it similar to over-drying DNA at the end of a protocol?

I dunno the science of it, but I know that too much of it definitely affects yield

 

[DNAdude120]

From Invitrogen:

​

​

Addition of too much chloroform: [FONT=Times New Roman,Times New Roman][FONT=Times New Roman,Times New Roman]If a larger amount of chloroform than needed was inadvertently added, add more TRIzol so that the ratio of 0.2 ml chloroform: 1 ml TRIzol is maintained. If too much chloroform is added, this will drive the DNA, and eventually the protein, into the aqueous phase.
..

 

[BlueElmo]

I'm sure the OP would've appreciated your sympathy if you had offered it two years ago

Holy crap, I made this thread back in Jan 2009. Feels like two lifetimes ago. Brings back memories. Thanks whoever brought this back up.🙂