Five more days

In five more days I am presenting my data to the department. It's not really that big of a deal as far as presentations go, but I don't want to embarrass myself in front of people I see on a daily basis. Not to mention, having a seminar looming has many positive side effects.

1. Extra motivation.

I try to keep myself moving at a steady pace in the lab, but despite my best efforts, sometimes motivation is lacking and I don't accomplish as much as I should. When a presentation is looming, it provides that extra little bit of motivation to get more accomplished.

2. New data.

Thanks to the extra time and effort put into lab work, I have an entirely new data set. This is not only great for my presentation, but it is also going to make my progress report for my post-doctoral fellowship that much more awesome. Of course, this data is all meant for a publication and I'm so close to wrapping up this paper I can almost taste it.

3. Analyzing Data

Putting a presentation together requires thinking about your data, something that can sometimes get overlooked when you are working your ass off. I often come up with new and interesting ideas when putting together a presentation and end up even more excited about my project that before. This is particularly nice since I am typically feeling a bit burned out as the seminar approaches due to aforementioned longer hours and quicker pace. It's like putting the seminar together is the cure for working your butt off in preparation for the seminar.

I didn't get everything accomplished...maybe 85%. The damn weather and crystal violet were determined to get in my way. While they did slow me down, I am still pretty happy with everything that I got accomplished.

Anyone else find motivation as a result of giving a presentation? If not, what does or are you one of those people that stay at optimal motivation all the time (a.k.a. people I am totally jealous of.)

Yep, it's been that kind of day.

I started the day off well enough. I woke up on time, I made myself some coffee (bonus) so that I wouldn't need to stop on the way. I get in the car and proceed to work, quite proud of the $1.50 I am saving. However, about 30 seconds later, the universe decided that today, I would provide it with comic relief or something.

Here is a break down of the events - and I'm just hitting the easy-to blog about items. Just imagine how my experiments went today.

1. Sweet ass (not the good kind).

While drinking my coffee on the way to work I realize that my coffee tastes awful. WTF? I ask myself. After another sip, I come to the conclusion that the giant container of half and half I just purchased isn't plain, it's french vanilla. GROSS. My coffee tastes like sweet ass!

2. My boob is really wet and warm.

I decide that my need for caffeine far exceeds the taste of the coffee so I continue to drink it. I'm having trouble drinking from the travel mug and decide there must be something wrong with the spout (or whatever that slide-open thingie is called). Oh well, slow coffee is better than no coffee. Right? Several labor-intensive sips later I notice that my left boob is feeling quite wet and warm. OH. MY. FUCKING. GOD. The coffee is leaking from under the lid and is now all down the front of my shirt and has apparently, soaked through my bra.

I consider going home to change, but cultures are waiting for me and my shirt is navy, so I ignore the signs of the shitty day to come and proceed to the lab.

3. Somewhere between 2X and 3X.

I arrive at my bench to find all the media that I prepared yesterday. The best thing I can say about the media is that it is sterile. Unfortunately, it also looks like it sat in the damn autoclave all night as the final volume is less than half of the starting volume.

I'm starting to feel a tantrum coming on, but I can find no one to take my rage irrationally out on at this point (lucky bastards) so I take a few trillion deep breaths and continue on with my day.

4. The sink stops working.

5. Fucking crystal violet!

I'm nearing the end of an adherence assay and start making the crystal violet solution. This go well until I try to filter the solution. That's when this happened:

Then I remembered that I was wearing relatively new shoes.

Five minutes into cleaning the mess.

Drink please.

Dumb F**king Donkeys Strike Back

I’m sure this is common in many labs, departments and offices: Some dumb fucking donkey (DFD) takes advantage of a good situation and everyone suffers because the people in charge prefer punishing everyone instead of confronting the person who is actually causing the problem. These instances might include, but are not limited to the following examples.

DFD situation #1:

DFD takes advantage of ordering by claiming that PI approved the purchase of an expensive reagent or kit (or whatever). After the item is ordered, it is discovered that permission was never granted and PI was completely out of the loop. This happens a few more times and involves the same person.

Solution: Enforce a new rule where ALL orders must gain approval from the PI, causing orders to get placed only once per week as PI does not have time to review orders multiple times a week.

DFD situation #2:

DFD refuses to turn in packing slips when an item arrives. This of course, pisses off the purchasing department, ultimately resulting in the lab manager getting bitched out.

Solution: Instead of speaking to DFD, the entire lab gets a 20 minute lecture on packing slips, what they look like, what they are for, why they are turned in to the purchasing department and what happens when they don’t. The tone of this lecture is extremely condescending and most lab members leave irritated because they don’t appreciate being spoken to like a child. As expected, DFD seems clueless and the problem continues.

DFD situation #3:

Instead of working a full day, the DFD comes in late and leaves early. Unfortunately, this DFD is the lab manager, so ordering is delayed; work orders for equipment repairs are put off, sterile items and media run out, etc. Lab members complain to PI.

Solution: Instead of asking the DFD to work more than 4.5 hours/day, the lab chores that DFD doesn’t want to do are redistributed to the lab members that actually work, i.e. post-docs and grad students.

Personally, I don’t understand why it is so difficult to confront the person who is causing the problem. I’m learning to take this shit in stride and file the experiences under, “Behavior I will not repeat when I obtain my own lab.”

Any DFDs in your lab/office/whatever

I can't say I'm too surprised.

I am not big on making resolutions. That's not to say that I don't make any, but they are usually the same and I tend to make them multiple times of year. Two of my on-going work resolutions are: (1) stay up to date on freezer stocks and (2) keep the lab notebook current.

I fail miserably at both of these on a regular bases because I hate making freezer stocks and am always putting off my lab notebook. In 2011, I've once again decided to make good on these resolutions. I've given myself the pep talk, the motivational speech, gone through the reasons why this is beneficial and how it would actually save me time in the long run, etc. If I was a wrestler or boxer, all that would remain was some screaming and hitting myself about the chest.

I wish I could report that this was my year, but sadly it doesn't appear so. I've really tried to make freezer stocks. I've grown some strains (3 or 4 times), but the cultures just end up sitting on my bench slowly dying until I decide to put them out of their misery and send them to the autoclave. This is the typical bottleneck in freezer stock-making.

The lab notebook is actually going a bit better. While I am not quite current, I am only @ 1.5 days behind. Not too shabby considering I've been as far behind as 6 months.

I'm not giving up. I am officially recommitting myself to the great freezer stock-making extravaganza of 2011. Let's hope I stay on the wagon.

Serenity Now!

Recently, I went from sharing an office with an insanely disgruntled grad student to sharing an office with an over-emotional post-doc (OEP) and all I can say is, I want the disgruntled grad student back.

OEP is working on a project that includes elements that are out of Magnum, PI’s (and the rest of the labs) expertise. However, I happen to know a shit-ton about this subject and as a result, I am the go-to person. In general, I don't mind this. I think part of working in a lab includes passing down knowledge and assisting your fellow lab mates. I draw the line at hand-holding.

At first, I tried to help. I explained concepts, techniques and went out of my way to make sure OEP knew where to look for information, and fielded numerous questions. This area was unfamiliar territory for OEP and I was happy to help. After a while, I expected OEP to take charge of the project and use me for troubleshooting or discussing data. I am willing to help, but the person I am helping needs to help themselves as well. Unfortunately, it seems like I am basically steering the project and I am wondering if OEP can function as an independent researcher.

I am constantly called on to look at data, explain data, explain what experiments to do next, etc. When I ask basic questions about results, I don’t get answers, I get blank stares. When I inquire about reading into certain topics I get answers like, “I didn’t know I should look at that.” When I ask, what are the limitations of this program or technique you are using, I get more blank stares. In a nutshell, OEP seems clueless.

Now, I would imagine that not having a clue is frustrating and I assume that this is why OEP breaks down in tears every other week. I am sympathetic, but to a point. I don't want to come to work and deal with someone crying or complaining about how they should just give up. It's exhausting and not to sound like a bitch, but I really don't have time for this. I've actually got my own projects to work on.

Am I being to hard on OEP? Maybe. When I started in this lab, I didn't know what the fuck I was doing, but I learned. I read many papers, I researched protocols, techniques, machines and brushed up on my biochemistry (big time). And yes, I asked my lab mates and Magnum, PI questions, but no one was holding my hand telling me what to do.

I'm at a loss as to what I should do. I don't even know that there is anything I can do. There really isn't anyone else in the lab that can help OEP. I don't want OEP to fail, but I am not willing to sacrifice half my time to steer another project and pick up the pieces every other week.

Week in review

In the lab:

Like everyone, my week in the lab featured it's fair share of ups and downs. Unfortunately, it also featured some stupid.

The ups:

Successfully electroporated notoriously difficult strain after being told it couldn't be done. (Yeah, don't tell me that if you want me to stop.)

Purified six proteins (of 12) for final replicate. This means my paper will likely be out by the end of the year.

The downs:

Cleaned up after one to many lab members and as a result, went off in lab meeting.

Dropped a bucket full of tubes containing turbid, smelly cultures. Of course, the tubes broke and there was glass and nastiness everywhere.

The stupid:

Performed FPLC without attaching the column to the machine. Doh!

It just might work this time \o/

The situation: I'm slated to present a poster (blech, I hate posters) at an upcoming meeting. When the meeting is two weeks away, the clouds part, the sun shines and all my cloning problems are resolved.

The question: Now that the constructs are complete, can I get the strains made in enough time to do experiments and include the results on the poster? (I've wanted these results on the poster all along, but after I stumbled into a bad cloning patch, I resigned myself to leaving this data off.)

The Answer: If I work everyday between now and the meeting, don't make any mistakes and find a way to print my poster at the 11th hour, I can include the data.

Sidebar: I consider throwing in the towel before I even attempt this whirlwind of tasks. You see, this shit happens all the time. I have a breakthrough within two weeks of an important meeting or presentation and bust my ass trying to acquire new data to include on a poster or in the presentation. All signs point to things not working out, but I try anyway and attempt to just push through only to finally realize a couple of days from the deadline that there is no possible way to get everything done. As a result, I am exhausted, irritated and consider my poster or presentation completely lacking as it doesn't contain the new data.

Current Standing: Well, I didn't throw in the towel. I am t-5 days from the conference. The strains are made and almost all the experiments are complete. By the end of today I should actually be in a place where I can put the poster together. If things work out, this will be the first time that last minute data gathering doesn't end in a complete and total cluster fuck.

The lambs are still screaming

Holy. Motherfucking. Crap.

I don't know if I mentioned this before; I am claustrophobic. As a result, I don't particularly care for small spaces like elevators, closets, etc. I tolerate them because I know that I can leave. However, if you want me to freak right the fuck out, just make it so that I cannot get out.

Today, I got stuck on the 37 degree room. I went in to put some cultures on a shaker and when I tried to exit, the door would not open. Immediately, I started to panic. I wildly yanked on the handle and pushed on that lever that is supposed to release the door. It didn't work. Suddenly, it smelled like a million liters of E. coli were growing and the temperature in the room spiked by about 10 degrees. I could feel my sanity slip, slip, slipping away.

Just before I started screaming and beating on the little window like an actual mental patient, I gave calm and collected one more try. I turned the handle and what do you know...the door opened. I got the hell out of there.

The incident is over, but I feel on edge.

I need a drink.

Seminars, meetings and clubs...just kill me now

I know that, at least in theory, journal clubs, group meetings, lab meetings and that fancy seminar given by a speaker from out of town are for my own scientific good. However, the fact that all of these things occur on a weekly basis is wearing me right the fuck out. My days are already a study in multi-tasking. I need to be productive. Just like everyone else I have deadlines and expectations to meet. Now that I have my own funding, I have progress reports that loom. I also have imaginary or self-created stresses, like worrying about my productivity post-child being compared to my productivity pre-child.

In the first month back at work, I know I wasn't all that productive, but I've seriously picked up the pace since then. I also want to prove to myself and any other nay-sayer, that I can still kick some scientific ass. As a result, my days are very busy and I get seriously annoyed when other work-related activities take me away from the bench. I feel like I need every minute. Seminars, meetings, whatever; they all cut into my day. Sure, they cut into it before, but now I can't just stay late. Of course, most of these things aren't optional, so I begrudgingly go and get nothing out of it. Why? Because I spend most of the time thinking about everything I need to be doing instead of listening and (hopefully) learning. Because of this, I get nothing out of these meetings and they truly do become a waste.

So what's a busy post-doc to do? Can I skip any of these meetings and not create a shit-storm for myself?

Lab meeting - For the most part, I don't actually mind lab meeting. I like to see what my co-workers are up to and provide (or receive) helpful suggestions. Besides, lab meeting takes place during lunch time, so at least I can multi-task my lunch.

Verdict: Skipping is not an option. Lab meetings are absolutely mandatory unless you are on vacation, sick, at a meeting or trapped under something heavy.

Group meeting - This is the departmental meeting. Sometimes people from other labs in the department present, sometimes people from other labs from local institutions present. Some of them are interesting, some of them are a snooze-fest. Like lab meetings, group meeting occurs at lunch so at least I can kill two birds with one stone.

Verdict: There are a lot of people that attend these meetings, so I'm not likely to be missed, but Magnum, PI attends them religiously, so missing too many is probably not an option.

Journal club: I fucking loath journal club, especially at post-doc university. The papers people choose are lame and I rarely ever get anything out of them. Plus, a dumb-fucking donkey from another department often attends these clubs and asks stupid and inane questions every five minutes. Also, they always run 10 to 15 minutes over.

Verdict: I think I can blow these off. Magnum, PI sees these more as required for students and I am not a student. Other post-docs in the department do not attend and nobody says boo about it.

Fancy seminar: To some degree, I think everyone should attend these. After all, I wouldn't want to give a talk at another institution and have only five people show up to listen. However, at least 50% of these talks are way, way, way out of my subject area. They are so far out that I would need to spend a week reading papers just to prepare to listen. Even then, I would still probably get lost half way through. This is because the speakers are invited by PI's from within the entire center and the center is composed of some very diverse departments. My department reminds of the sesame street song, "One of these things are not like the other."

Verdict: I think I will make an executive decision to only attend the seminars that I find (a) interesting or (b) are at least tangentially related to my area of research.

Final verdict: Attend lab meeting and the departmental meeting every week, attend the fancy seminars when I am so moved and give journal club the finger. This sounds much, much better.

What do you guys do? Do you attend every seminar or do you have the luxury of picking and choosing?

Spread the love around

Dear new lab person who shares my office,

I am really glad you joined the lab. You seem smart, funny and motivated and I really love your accent. However, it would be totally awesome if you could just sometimes, like once a day, ask someone else in the lab for help. I know for a fact that they know where all the media, tubes, glassware and equipment is located and I think they will at least point you in the general direction. And pretty please, with sugar on top, stop lingering behind me when I am working at the bench. This does not make me work faster, not to mention it is distracting me and making me less-inclined to want to help you. Maybe I am just weird, but I really don't like it.

Sincerely,

MXX

WTF did you just call me?

I was reading a post over at Profgrrrrl's and it got me thinking about names. As I mentioned in my last real post, there is a person employed here who does not bother learning the names of the people who work in the lab. This might be because they (a) are too lazy (b) have difficulty remembering and pronouncing foreign names or (c) are just plain rude. Personally, I think it is a combination of a and b. Because s/he doesn't know our actual names, s/he must come up with other things to call us. Examples include:

Skinny guy - a.k.a. my graduate student. He is Asian and yes, he is skinny, but not overly so.

Hat guy - this is the M.D./Ph.D. student in the lab that wears a baseball hat almost every day.

Tall girl - M.D. working in the lab.

The best/most annoying one (depending on your perspective) is:

Canteen. This is how s/he refers to me. S/he tried to read my name off of a shipping label and this is what s/he came up with. Now, my name is a bit difficult to pronounce when you read it. People typically say it one of three ways, none of which are correct. I thought I had heard it all, but apparently I has not.

Here is what Canteen has in common with my real name:

(1) starts with a C

(2) contains the letters N and A

(3) is two syllables.

Here is what Canteen does not have in common with my real name:

(1) My name is not Canteen

(2) My name does not have "teen" or anything that looks like "teen" anywhere in it.

(3) In fact, my name does not contain the letters T or E

(4) My name contains 4 letters total.

Nope, I don't have the ability to communicate with ESP.

After working here in post-doc lab for just over a year and a half, I've noticed a thing or two about how the lab is run. One of those things is that my mentor, Magnum, PI is a total puss about letting people go. As a result, there are a few individuals who, on a good day, qualify as useless. One such individual is in charge of (1) ordering, (2) sterilizing and (3) keeping certain lab items stocked (i.e. 50 ml conical tubes, sterile pipettes, tips of all shapes and sizes, 96-well plates, etc.) Because this person has a zeal for autoclaving, and nothing else, I will refer to them as the sterilizer (TS). TS does not exhibit any inclination for ordering or keeping items stocked. This causes me and many others in the lab to make frequent trips around the floor tracking down tips, tubes and missing orders.

Before I left on vacation, we were completely out of 200ul tips. TOTALLY FUCKING OUT. Like a great many other researchers, I use 200ul tips all day, everyday. These are essential and I get very upset when I do not have any. I get even more upset when I subsequently find that we are also out of conical tubes, small gloves and 5ml pipettes. Thankfully, I went on vacation before I located TS and punched them in their idiot face. Over the next week my rage and desire to punch idiot faces subsided and I returned to work to find everything restocked, except small gloves. I felt a sting of irritation when I noted this, but decided not to dwell. However, this all changed today.

Note: Before I left for vacation, I ordered a chemical. Suffice it to say, that if I order a fucking chemical it is because I need it, not because it comes in a pretty bottle or because I am trying to round out my collection of chemicals that start with beta or whatever the fuck. Yesterday, this chemical had not arrived. Of course, TS wasn't here, so I had to wait until today to have this glorious conversation and trip around the floor.

MXX: "TS could you please check on a chemical that I ordered two weeks ago. It really should have come in by now."

TS: "You haven't received it?"

MXX: "Noooo." (In my mind I am saying: NO YOU FUCKING IDIOT! Why do you think I am asking about it??!?!?!)

TS looks through orders and finds the order form I handed in, then informs me: "It came in, PersonX put it away."

MXX: "Thanks." I proceed to person X

MXX: "TS told me that you received a chemical that I ordered last week. Could you tell me where it's at?"

PersonX: "I gave it to that skinny guy in your lab." (note: that skinny guy is what person X calls my graduate student as person X doesn't bother to learn our names.)

MXX: "OK. Thanks." I proceed to skinny guy and ask him about the chemical, but he informs me the only thing he received for me was ligase, which I already knew about. So, I proceed back to TS.

MXX: "Soooooooooo. Person X claims they gave it to my grad. student. My grad. student only received ligase, not the chemical I am looking for."

TS (looking at packing slip again): "Let me call the company, it looks like the chemical didn't ship."

MXX: "uhhhhh, OK" (In my head: What fucking new magical information just appeared on the sheet that now explains it didn't ship, when 10 minutes ago, not only did it ship, but it was noted that a specific person put it away????!!!!!!!)

TS calls company and then turns to me: "Oh, that's right. The order was cancelled because the item was discontinued. Remember, I told you that."

MXX: "Really? When exactly did you tell me that?"

TS: "Last week."

MXX: "You mean, last week when I was on a rock in the middle of the pacific ocean?"

TS: "Yes."

MXX: "Seriously? Was this a psychic message or what? I WAS NOT here last week."

TS: "Hmph. Well, I know I told you."

MXX: : LOUD sigh. "Whatever!" (In my head: Must not kill useless idiot. Must not kill useless idiot. Must not kill useless idiot.)

I'm not the boss of you

There are a couple of graduate students who feel the need to make sure that I know how many hours they spend in the lab. First of all, I don't care how much time these people do or do not spend in the lab because:

(1) it's not my job

My job as a post-doctoral fellow is not too police how much or little graduate students work. I think that my job (right this minute) entails (a) conducting research, publishing papers, obtaining funding and acquiring skills that will assist in obtaining a tenure-track faculty position; (b) mentoring my little grad student with stinky breath, (c) acting as a sounding board for new ideas and troubleshooting for any lab member, (d) editing grants and other written items for my Magnum, PI and (e) random lab shit. I'm sure I've left something out, but you get the idea.

(2) it's really more about the quality and quantity of data you produce.

It's all about the data. As long as you are producing data at a reasonable pace and publishing papers, all is well. If you can do that working 3 hours a day, great. If it takes you ten hours a day, fine. I really don't care.

(3) it's not my lab.

I have no authority to reprimand someone if they choose to do nothing or not come to the lab.

And now for the point of this post:

I've started getting to the lab about an hour and a half earlier than what I used to as my child likes to get up at the crack of ass. Since this time, I've noticed that the people in my lab that were always talking about how they got to the lab by 6:00 a.m. NEVER get here by 6:00 a.m. In fact, it's more like 8:00 a.m. Maybe it's a coincidence. The same thing happened in grad. lab. There was one fellow grad student who whined endlessly about how they never could sleep in and were always in the lab by 5:30 or 6:00 a.m. Low and behold, when I start getting to work by 8:00 a.m. do to a schedule change, their sleeping problems suddenly vanished.

Like I said, I don't give a flying fuck when these people get to work, when they leave work or what they do while they are at work. I'm just surprised that people regularly take the time to lie about this shit.

Busy with a short fuse

It's been nose to the grindstone time for the last couple of weeks. Unfortunately, that's not really producing more data. It is, however, creating one tired and cranky microbiologist.

At this point, I am in the midst of troubleshooting. As we know, troubleshooting does not equal data. Therefore, I working more than 40 hours a week. Normally, this is not a problem. However, with less than two weeks to go until my due date, it's tiring. The internal battle isn't doing me any favors at the moment either.

Tired, pregnant me: "Fuck this shit. Just try to get done whatever you can within the amount of time that you have and let it go. It will be here when you get back"

Career-driven, microbiologist me: "Don't be such a whiny little bitch. If you don't get this stuff accomplished, you will be kicking yourself later. If you can be in a position to do some serious writing, you will have something you can work on from home and your transition back to work will go much more smoothly. Besides, if you try really hard and still don't get this finished then you'll dwell on it less as you know you did the best you could do."

Career-driven me is still wining, but tired pregnant me is making serious gains.

Lab coat fail

I can't really blame my lab coat. (Picture is taken from my perspective.)

Argh.

Sometimes I think the universe is trying to determine just how far I can be pushed before I completely lose my mind. I can tell you right now, it’s not that far. In general, my fuse is pretty damn short. Patience is not a virtue I possess and I do not fear or avoid confrontation. However, I really do try to keep myself under control, especially at work. If you compare myself of today with myself of 10 years ago, you would find significant improvements in this arena, but if you compared me to myself of just one year ago, you would probably find little improvement.

Then there are weeks like this, where you might find it difficult to see any progress no matter which version of myself you compared me with. It feels like 10 million little things are going wrong. No, none of them are worth losing my mind over, but as they add up, I find myself dangerously close to the breaking point.

As an example, let me describe a 1-hour portion of my day yesterday:

• Our lab possesses multiple thermocyclers and right now, we are down to one. I think fixing one of them would be nice, you know, just in case that last one decides to die. Yesterday, the last thermocycler died. Fortunately, it was some sort of issue with a fuse and was repaired quickly, but considering I cannot do what I need to do without PCR, I was pretty damn irritated.
• I send DNA for sequencing. I get results using one primer, but no results using the other primer. I use the sequencing facility's primers. If it were my DNA, then neither primer should have yielded a result. Therefore, I think it is their primer and that they should re-run the samples for free. They are not under that impression.
• I open the -80 C and a mountain of tubes fall all over the floor. Someone decided that balancing an open box precariously near the front of the freezer was a dandy idea. I want to leave their stupid shit all over the floor, but I pick them up and shove them back into the freezer.
• After picking up all the tubes and replacing them in the freezer, I try an locate the strain I need, only to find that whoever got into the stocks previously completely rearranged all the boxes and since they are covered in frost, I have to remove them one by one, open them and pull out a vial to determine which box I am in. No, I don’t fix the order, because I am annoyed as shit.

This kind of irritating crap has happened every day this week. I am so ready for the weekend. I need to rest and reboot. I really hope next week is significantly better.

Sure, why not?

Magnum, P.I. informed me today that I was presenting at out next data club. This is something that we do with a few other lab. It is more work than lab meeting, in that it is an actual talk, but it is still considered informal.

I actually like presenting and getting feed back, but I'm not exactly bursting with free time inside or outside the lab right now and I'll need to prepare a lot of new slides since I haven't presented most of this data before.

Really though, what's one more thing? and Hey, at least I got to choose the date. :)

Would you like me to yell at you?

If you find a centrifuge in another lab with a sign on it that reads, "Out of Order. DO NOT USE!" Would you:

(a) Find an alternative, knowing that using a broken centrifuge can damage the centrifuge further and even cause severe destruction or injury.

(b) Ignore the note and use the centrifuge anyway.

(c) When busted using the centrifuge, argue with the person who catches you, citing that since the centrifuge could be turned on, it must be functional.

(d) Continue to argue with person, even after they explain to you that the error code indicates that you should cease using the centrifuge immediately and contact the manufacturer.

If you answered b, c or d, then I can only conclude that you want me to yell at you. You may also conclude that you are a jack-ass.

Cloning tard

That's me, at least for the last couple of days. Typically, I am a cloning rock star, but a combination of arrogance and laziness resulted in a 1 day process taking 3 days.

Here is the situation.

Currently, I'm in the midst of a protein trouble-shooting extravaganza and as a result my work load is significantly lighter than usual. To fill the gaps, I decided to pick up some cloning that I abandoned this summer.

2 days ago:

After much digging around in the freezer, I located the five abandoned PCR products, ligated them into a vector and transformed the ligation.

That's pretty standard. I left feeling confident that I would find five plates containing beautiful colonies the next morning.

1 day ago:

Success! The perfect amount of colonies were scattered about each plate. You know, not so many that you can 't pick a single colony, but not so few as too indicate that all you probably got back was a bunch of bullshit re-ligated vector.

Anyway, I colony PCR a few colonies off of each plate and run a gel, only to find that I didn't get a single PCR product! WTF? This doesn't happen...at least not to me. I stair at the computer screen in disbelief for a few seconds when I realize that in addition to a lack of positive results, I also lacked a single negative result. I should have one or the other.

I dig back into the recesses of my mind and vaguely remember that the inserts in this case would be pretty long. Most likely, I didn't use a long enough extension time.

I set up another colony PCR, this time with a longer extension. I run this PCR out on a gel and this time I see a couple of PCR products, but most of the gel is black. Seriously, WTF? A couple? What am I? A first year grad. student.

At this point, it's the end of the day and I really want to get the hell out of there, but I am not willing to admit defeat, so I decide to inoculate a couple of colonies from each plate.

Today:

I verify the long way (the short way being colony PCR the day after you transform). I mini-prep the overnight cultures, digest the plasmids and run a gel. The image of the digest comes up on the screen and it's all I can do not to yell out MOTHERFUCKING FUCK! It looks like all empty vector, except for the two that I already knew about from the day before. I walk down the hall in a huff toward my office.

I'm ready to trash three of the plates, but instead I decide that maybe, just maybe it's time I actually look at a gel from the summer and see exactly what damn size the PCR products I dug out of the freezer were in the first place.

Well, what do you fucking know? Three (that's right three) of the PCR products are exactly the same size as the vector I ligated them into. Damn it. The digestion was a complete waste of time.

I don't feel like digesting again with an additional enzyme because I don't know which one to use and at this point I am not in the mood to look it up (despite the fact that not looking shit up in the first place is exactly what got me into this mess). I decide once again on PCR, but this time I will actually use the correct extension time.

Conclusion, almost every vector contained the appropriate insert.

I could have found this out yesterday if I had just looked up the gel in the first fucking place and used the correct extension time for the first round of colony PCR. Now, I'm behind, I feel like an idiot and it's only Monday.

Snow, sleet and pink gloves

As I drove myself into work today I witnessed an event that rarely occurs in my home city; rain, sleet and snow all coming down simultaneously. In addition, my umbrella was safely tucked away in my office, far across the parking lot from where I exited my car.

In other news, me and eight other science-types made comments about a friends Facebook status relating her recent and exciting acquisition of pink latex gloves. One of the final comments, summed the whole exchange up quite nicely. It read, "It's kinda sad what scientists get excited about."

I found this funny and true.