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'd like to shove a tree up Santa's butt

Seriously folks, my holiday cheer is all tapped out. I'm pretty sure I ran out a couple years ago. Don't get me wrong, I like the days off, but I dislike, dare I even say hate, almost everything else related to this holiday and since I can't drink or beat the crap out of the next person I see wearing a holiday sweater, I am going to use this blog as a place to let out my holiday-induced irritation over the next few posts.

Topic 1: Step away from the holiday music. For reals.

I don't want to hear stupid holiday- themed music on the radio station, in stores, at work or anywhere else anymore. Every damn store in the city started playing this crap in early November* and every song has already been played a trillion times. What's worse than just hearing plain old x-mas tunes over and over again, is that there is a limited number of songs from the start, so instead of just playing all of them ample times and retiring them, you get to hear every other musician's take on the same x-mas "classic" and it always really bad, even if the artists normal music is good.

My mother contributes to this epidemic of shitty music in her own special way. She loves her x-mas music and as a result, we must listen to it on Christmas day, while opening gifts. One of her favorite holiday CDs is a Reba McEntire does the holiday classics type of deal. To say it's one of the worst things I've ever had the misfortune of listening to, is an understatement.** When I hear her voice singing x-mas songs, I feel homicidal and hope for spontaneous deafness that lasts around 45 minutes. If we are lucky, we get her to play something like Bing Crosby does Christmas (or whatever his particular album is called), which isn't nearly as bad since at least he sang all the songs without trying to add some stupid unique flair. Hell, I probably wouldn't even mind listening to ole Bing on x-mas day if I hadn't already been listening to bullshit holiday music for the last 60 fucking days.

For my own well-being, I think I'll just keep the ear buds in as much as possible until the holiday season is over.

*Some stores were already playing this crap in October. October!!

**In all fairness, I must admit that I do not like country music, nor do I enjoy Reba McEntire. However, I love the hell out of Radiohead, but I don't want to hear their version of Oh Holy Night either.

An open letter to the I.T. department

Dear I.T. staff,

You dumb fucking donkeys are completely useless. I glimpsed this on my first day at post-doc institution when you walked into my office, hung your head, sighed and said in a defeated voice, “Great. You use a Mac.” Remember, I connected my computer to the network printer while you watched. I assumed your ineptitude was Mac-specific, but today I realized that it’s likely technology-specific.

I'm not going to lie. It’s disconcerting to find out that the network at this university is infected with a virus, but it's totally fucked up, to learn that the network became infected with this virus over three weeks ago and no one informed the people who hook their computers up to said network until today. Apparently, you didn't think it was a big deal a few weeks ago because only a few computers were infected.

Do you jackasses even know what a goddamn computer virus is? I think you don’t, and since your unlikely to get fired, I feel the need to at least try and educate you.

A computer virus is a computer program that can copy itself and infect the computer without the permission or knowledge of the computer operator. Furthermore, the virus can spread to other computers via floppy disk, CD, DVD, USB drive and networks.

Still confused?

Computer programs are instructions for a computer. A computer is a machine that manipulates data according to a set of instructions. A machine is any device that uses energy to perform an activity.

Regards,

Microbiologist XX

Bullf*ckingsh*t

I was completely taken aback today when the following University of Alabama thesis, written by Jacksonville State University president, William Meehan, was brought to my attention. (as seen on Orange Crate Art). What the fucking fuck people? 

And it gets better, according to various sources (here, here and here).

Apparently, this recent finding stems from a civil law suit filed in 2003 by Jacksonville State professor R. David Whetstone, who claims Meehan took about 55,000 plant samples. The dissertation is supposed to serve as an example of how Meehan had stolen other's work in the past.

But that's not all,

In 2007, Meehan was busted using copy from the internet in his "Town and Gown" newspaper column. At that point, Meehen claimed that the articles were ghost written by a former university publicist even though the articles clearly contained his name in the by line. 

And this is what really fucking pisses me off:

The University of Alabama reviewed the matter and was "not inclined to go behind the decision the dissertation committee made 10 years ago in granting Meehan a doctorate in education, said spokeswoman Cathy Andreen in a statement Tuesday. She would not elaborate. 

Alabama's position closes the matter for Jacksonville State, said JSU board chairman Jim Bennett. "We're satisfied that President Meehan has a legitimate degree," said Bennett, who also is Alabama's labor commissioner.

Excuse me Jim Bennet, but a legitimate doctorate requires a legitimate dissertation, and a legitimate dissertation cannot be plagiarized. In case your wondering, a document that is not plagiarized does not contain sections that are word-for-word identical to a previously published document. 

Fucking donkeys.

Damn these sequences to hell I say

I just received THE WORST FUCKING sequencing results EVAH! 

First of all, I fucking HATE dealing with sequencing. It's tedious, it's boring and it takes time away from actual bench work.

At my old institution all the sequencing was done at a core facility and rarely did I get anything back that wasn't pristine. There is not core facility at my new institution, so I am using a new company and I am NOT impressed.

Sure, it could be the plasmids I sent them, but I seriously doubt it. Here's why:

I checked the concentration and purity before sending the plasmids off and they were all very good.

I am using vector-specific primers that are provided by the sequencing facility and I get the following results:

Primer 1:

gives sequence results ranging from beautiful to tolerable.

Primer 2: (same template as primer 1)

gives sequence ranging from 300 nt reads (300?!?!?) to 800 N's with a few random A's T's C's and G's dispersed throughout. 

Good and bad sequence from each primer do not correlate with the template. If the worst sequence from each primer came from the same template, then I would assume the template that was the problem, but considering it did not correlate... 

I'm taking a break before I blow a gasket.

InaDWriMo 2008 progress report and other fun stuff

Well, according to my status bar, I am 40% of the way to my projected goal for InaDWriMo. However, progress is temporarily halted since my boss wants me to stop writing. I told her OK, but I am writing in my free time so I think I'll stop at the end of the month.

Now for the other (not so) fun stuff.
I am actually in a shitty mood because things aren't working. There is a month remaining in gradstudent lab and I need to finish more than a months worth of stuff. When I defended my dissertation in September, I thought that there was ample time, but that was before the triple mutant from hell.
I need to make a triple mutant lacking genes A, B and C. Right now, I have double mutant AB and single mutant C. I've tried introducing the knock-out construct for deleting genes AB into mutant C, but it is toxic and the cells get really pissed and lose the KO construct before homologous recombination can take place. I tried to get around this by growing the cells in the presence of antibiotic since the vector contains resistance to one antibiotic and the genes are being replaced with a cassette that imparts resistance to another antibiotic. I obtained many colonies with the correct antibiotic resistance phenotype, but all of them still contain the genes I am trying to delete. The number of colonies and the rate of growth of said colonies indicate that they are not spontaneous, antibiotic-resistant mutants, indicating that the resistance gene is somewhere in the chromosome. Where? I don't know. I only know where it is NOT.

Obviously that wasn't working out, so I tried the opposite approach. Delete C in the AB double mutant background. Fine. I introduce the KO construct and the cell grow normally (yay) Unfortunately, that is the end to the good news. The KO vector for C is a temperature-sensitive vector and gene C is only deleted, not replaced with an antibiotic-resistance gene or cassette. So, it is screening hell. This approach works in the following way: The vector is integrated after growth at a non-permissive temperature. Cells are returned to the permissive temperature and and are passaged several times. At some point dilutions are plated on media containing no antibiotic. These colonies are replica plated onto additional plates that do contain antibiotic. Those that are sensitive, are screened for the mutation using PCR...colony PCR.
Unfortunately, the locus I am deleting doesn't seem to amplify using colony PCR, so I ended up trouble-shooting PCR. PC fucking R people. This is like trouble-shooting shampooing my hair. Finally, I found primers that work, except they don't yield a product if the gene is deleted. Awesome. A fucking negative result. This means that I have to recheck any colonies that don't yield a product and so far, all of those potential candidates have yielded a product the second time through. Still, it is better than growing up 5 zillion cultures and extracting DNA for PCR. Now, I am adding an antibiotic-resistance gene to this construct, so that I will have a selection. Good thing I am a rock star at cloning.
I am also considering that this triple mutant is lethal, although it really shouldn't be. We'll see. I am going to give it the ol' college try one last time with the new vector and see how it goes. If I can't get this mutant, then I will just have to drop the strain from the manuscript. :(

So, yeah. I am irritated to say the least.

Just open the damn door already.

An open letter to the lazy people who work in my building:

Please, please, for the love of god, open the doors to the building using your actual hands and arms. It is really easy, just grasp the door handle, pull toward you (unless otherwise noted on the door) and walk through. Unless you are disabled or have your hands full, you don't need to press the button to open the door.

First of all, it actually takes more time to use the button than open the door yourself. You have to press the button, stand there (while blocking the path for everyone else) and wait. It also causes delays for everyone else because even though the button opens two doors, these people seem to enjoy standing right in the middle, so that when both doors open just enough, they can squeeze through. Meanwhile, everyone else is standing there, waiting to get in.

Second, it is fucking wasteful. Even though it only takes an able body person, carrying nothing, a couple of seconds to walk through the doorway, the doors remain open for significantly longer. So, long after the lazy person who couldn't be bothered to use their arms to pull on a door have walked away, the doors are still wide open while cold air billows out. Sure, the school foots the electric bill, but it is inexcusable to be this wasteful in order to avoid opening a damn door. If you don't know what the big deal is, then you need to pull your head out of your ass.

Finally, stop taking the use of your arms and legs for granted. Those buttons are there for people who cannot open the door without assistance. Almost everyday, I watch a man who drives his wheelchair around with a barely-functional finger use those doors. Do you think he wouldn't rather open the door like everyone else? When you roll up in a wheelchair, you too can use the button. Heck, if you walk up to the door carrying a ton of crap, you can use the button, but everyone else, burn the extra 2 calories required to manually open the door yourself.

Regards,
Microbiologist xx