Dedication or Stupidity?

It is the week of Christmas.

The weather has turned seven shades of ridiculous.

The boss is out of town for the week.

My entire lab is here working (except for, you know, the boss).

And, just on a personal level, my husband is in town and I still don’t have all of my Christmas shopping done.

What the hell are we doing here?

———-

Thesis:

So, my advisor already made CPP’s suggestion of doing all of the figures and figure legends first.  However, after spending hours of time moving images miniscule distances to the right or left and changing the font on the figures several times, I had to accept that I was procrastinating.  So, I thought I should start writing the text of the Results.  I still have a couple of figure legends I could write, though.  Maybe I should do that.  Especially since all I’ve really done is change forms of procrastination.

I’m not even thinking about the introduction.  There is no introduction in my fantasy world.  Panic does not even come close to describing how I feel about the introduction.  I’m just going to get through the results first.

———

Sleep:

I’ve got the sleeping pills.  They helped me get to sleep, but I didn’t stay asleep for the first couple of nights I took it.   Then, I started sleeping through the night and needing to sleep through half of the day, too.

So:

No sleeping pills = no sleep, tired all the time

+ sleeping pills = lots of sleep, sleepy all the time

Not sure this is an improvement.  I think I’m going to try a different sleeping pill and see if that’s any better.

———

Finally:

If someone could make my super simple subcloning work right, I’d appreciate it.  I really don’t understand what the problem is.   Cut fragment out of vector 1, ligate into vector 2, transform, miniprep.  But, then I do a digest to check if it’s right and the digest comes out all wrong.  On all of them.  This is the second time I done this damn cloning.  If I have to do it a third time, I’m going to be wicked pissed.

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3 thoughts on “Dedication or Stupidity?

  1. This is the order in which I usually write papers (and how I wrote each chapter/paper for my thesis):
    1. Methods (easy to write them as you do them)
    2. Figures/tables
    3. Results
    4. Discussion
    5. Introduction
    6. Abstract

  2. i agree with PIT, i do the intro last…it causes hyperventilation 🙂

    as for the subcloning, if your restriction digest is wrong, is it possible the plasmid is not what you think it is? we’ve had that prob in the past as we generally pass vectors around to keep from having to buy them. if you haven’t, maybe digest the empty vector to verify it’s right, then digest sequentially (if it’s directional cloning) before ligating. another prob i had was enzymes that were older and decided not to cut supercoiled plasmid really well and FUBAR’d my cloning for a month or so (i was slow to pick up on this).

    good luck 🙂

  3. A few things with the cloning:
    -Sequence your vector. Seriously. Especially if you got this from someone else (commercial or otherwise).
    -As the above poster suggested try doing the digests separately.
    -Check the DTT in your ligase buffer. If it doesn’t smell bad, then it’s bad. (I’ve had a problem with getting a bad batch of buffer from NEB.)

    If you’re still having problems, send me an email. I do metric-crap tons of cloning. (And have troubleshot just about everything)

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