A Rant about Western Blots

I hate Western blots.  I think they are the most annoying, frustrating, and obnoxious experiments to do in lab.  I hate that the process takes at least two days and that it's so antibody-dependent.

First, I am fully willing to admit that I am not good at Western blotting.  My films usually look a little messy, and I've taken several steps to clean up my methods to no avail.  The process is so touchy and there are so many steps that, apparently, I have trouble performing properly.  That's my problem though.

What I hate about Westerns is how subjective they are.  One antibody uses a milk block; another uses BSA; the last one takes three quarts of finely grated basil and a dash of kosher salt.  The first one needs to be incubated overnight at 4 degrees; the second needs three hours at room temperature; the third needs to be set on a wooden stool in the midday sun from exactly noon to 2:30 PM.

The "voodoo" science involved in making these antibodies work bothers me.

It also doesn't help that the department's developer was made prior to the Cold War era (and I'm probably not even exaggerating too much).

What alternative is there?  None.  I may hate the process, but there's no way I'm getting around doing Western blots for the rest of my laboratory career.

Science music!

I like listening to music a lot, and I always find it comforting to listen to it in lab.  This often leads to me basically choosing what everyone else has to hear... which was fine when there were two people in the lab.  Now that we've grown in size, however, I feel bad about making everyone listen to my music.

In reality, they have every right to plug their iPods into the stereo or choose a different radio station... but they don't.  So I do.  Does this make me a bad person... because I listen to my music all the time in the lab?  Maybe I would feel bad about it if anyone else expressed interest in playing their music.

I guess that I'll just continue listening to Britney Spears and the Spice Girls on a regular basis.  Nobody's complained yet...

My lab playlist:

• Joss Stone
• Girls Aloud
• Maroon5
• Clarika
• Britney
• The Saturdays
• Janelle Monae
• Amy Winehouse
• Nicki Minaj
• Ke$ha
• Erykah Badu
• MGMT
• Paula Abdul
• Spice Girls
• Alicia Keys
• TLC

I'm not entirely sure why they're almost all female singers, but I guess that's my preference.  I like to think that I have a mixture of music, but I tend toward radio-friendly pop music.

Pressing the "Submit" button

As I mentioned in the last post I did, my PI is gone on vacation and I've been working my butt off with a number of different experiments.  Today, I finish the last experiment we need to submit our latest paper.  The problem is that she's out of the country... leaving me to submit the paper.

She sure didn't seem to have any problems with me submitting the paper.  Maybe I should take that as a sign that she trusts me to do this on my own...  But it's still freaking me out.  What if I upload the wrong file?  Did I choose the right keywords?  Is my new figure "pretty" enough?  I shouldn't be worried since it's not an enormous paper, and it's not going to a super-high impact journal.  Still, it's my first time...

Submitting this paper is scary and fun at the same time.  I guess it's a good experience for the future, regardless of how I feel about it.

While the boss is away...

My PI is on vacation to a far-away land.  And I am not.

As usual, when the boss is away, everyone in the lab is even more stressed than ever.  I'm still trying to finish up the comments on the paper we submitted last month, so I'm under a deadline to submit it before vacation's end.  Everything would have been done last week had I properly read the labels on our antibodies.

Dammit.

But all is not lost - I will finish my experiments this week, get back on track, submit the paper, and maybe even relax ever so slightly.

On a positive note: the experiment I so desperately needed to work for my thesis actually worked.  The universe doesn't hate me!  And the best thing is that I included almost every single possible control in my first set of experiments.  Now all of my data is organized, beautiful, and figure-ready.  I've never had my experiments work so beautifully on the first go.

I could get used to that.

Oh, and I changed the background on the blog because I was playing with Illustrator and liked what I made. :)

No time for sickness

The Independence Day weekend was fantastic, visiting my undergraduate school and bumming around town during the hot, sunny, beautiful summer days.  At times I miss being an undergraduate, particularly in the summer, when I had few responsibilities and I had a lot of free time to just relax.  Those days are apparently over, and while I might reminisce, I feel that it is truly time to move on and be a "responsible" and "mature" graduate student.

And while on that mini-vacation, I became sick.  It started just as a sore throat, and now it's going into full-blown sinus nastiness.  I can't breathe and my head feels like it's stuff in a giant feather pillow.  It's not like I'm dying, but I'm not enjoying this current state either.

Regardless, I don't really feel like working or thinking today: my head is tired, I feel sick, and my vacation days just ended.  I would like to crawl into a small, science-less ball and take a nap.  It's not like I'm asking for much.

But no. Cells must be split, infections must be inoculated, plasmids must be digested, and primers must be ordered.

I'm just going to do it all slowly.