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.)

Get back to where you once belonged

That is the theme of this week. Actually, my theme for this week is start the process of getting back to where you once belonged. This is the first week that I actually feel like a human. I'm adjusting to the sleep deprivation and mini-xx is getting into a routine which is allowing me to start accomplishing some non-baby related activities.

Work.
Although I didn't get all the experiments completed that I wanted before I began maternity leave, I did acquire enough data to start writing my first manuscript from post-doc lab. Since I want to get a good draft together before I return to work in a few weeks, I decided that I must get started writing this week. While I am happy to report that I did start writing, I am less thrilled about the progress I am making. Monday and Tuesday were down right painful. I found it difficult to concentrate and I could not get into a groove at all. On Wednesday, things weren't quite as painful and I actually managed to crank out an entire results section.

Appearance.
I also decided that this was a great week to start exercising again. After all, it's been two and a half weeks since the baby was born and three and a half weeks since I exercised at all. Not to mention, my best friend is getting married in Hawaii in August and I am in her wedding, which means I'll be in a ton of wedding pictures and I'll need to feel comfortable in a bathing suit. I wanted to start slow, so on Monday I started walking 2 miles a day. Next week I will increase the distance and start adding in some jogging. (I actually tried jogging this week, but my old sports bras don't really accommodate my near porno-sized boobs, so it was a bit painful.)

I'm keeping my fingers crossed that I find my groove by next week.

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.

Productive, pregnant, postdoc - Post 2

As I stated in my last post of this nature, my goal for this week was to combine all three PCR products and get them ligated into the first vector, pGEMTeasy. In short, I failed. However, I did get quite a bit accomplished. For ease, I've posted the picture from last week. (Come on, you know you love it.)

As far as the PCR goes, I completed PCR round three for 20 of the 26.

Of those 20, 13 of the PCR products were successfully ligated into the first cloning vector and are getting sent off for sequencing today. (I am dreading looking at all this sequencing data. I freaking hate it! It bores me to tears.) I will ligate the remaining 7 PCR products (which I just obtained overnight) over the weekend and transform them on Monday.

So, in all, not quite where I wanted to be, but not exactly too far behind either.

My goal for the end of next week is to get the final 6 PCR products created (i.e. combine segments 1,2 and 3) and ligated into the first cloning vector. Obviously, everything needs to get sequenced. Finally, I hope to sub-clone at least half (that's 13) of the pGEMTeasy constructs into the final vector.

Productive, pregnant, postdoc - Post I

My major goal at work right now is to get myself to a point where I can work on two manuscripts during maternity leave, IF it’s possible. To do this, I need to get a certain number of things accomplished. Unless the baby decides to make a very early appearance, I think I can get all of the following items accomplished in the next 7-8 weeks. I will post on my progress each week.

Manuscript A: This is a manuscript from graduate school that I started writing in June. I need to verify some in silico promoter analyses using semi-quantitative RT-PCR. To complete this task I will need to do the following, in triplicate:

• Order primers. (not in triplicate :))
• Grow strain under inducing and non-inducing conditions.
• Harvest cells at a specific time point and extract RNA.
• DNAse-treat samples, quantitate and perform RT-PCR

Not too bad, really. The only hold up here is an issue with strain availability, and this should get resolved shortly.

Manuscript B: This is my first paper from my post-doc. Originally I thought we might get this paper out near the end of last year, but I decided to add some experiments and broaden the scope of the paper. These extras are exactly what I need to accomplish.

I need to make two sets of 13 strains (yes, that’s 26 strains) for some in vitro and in vivo work.

Currently, I am cloning as I must create 26 constructs. For each construct, I must combine 3 PCR products. So far, I am about half way through this PCR nightmare. I’ve included the cartoon I put together to help all the non-cloners in my lab understand what I am doing. (Yes, there are some errors in my math regarding the cycles, just ignore it if you can.)

My short-term goal for the end of this week/early next week is to get all PCR products combined (resulting in 26 final products) and ligated into the first cloning vector.

Here we go again...

In yet another attempt to eat better and live better, Lancelot and I are making changes.

Change 1: get off our asses.

This change actually started a few weeks ago and so far, it's going well. We try to walk and/or jog for a distance of around two miles, two to three times per week. My personal goal is to get up to 4 miles, three times a week. I can make it just beyond two and Lancelot can make it just under two. I don't know what Lancelot's overall goals are, but the fact that he is even going is good enough for me.

Change 2: eat better

This is actually quite difficult as Lancelot and I eat out a lot. When we do eat at home, we typically eat frozen something. To transition into eating healthy without making ourselves so miserable that we want to quit, we needed to break this change down into baby steps.

baby step 1: eat at home

There is no requirement that the food is healthy and the only goal is to get adjusted to cooking and eating at home. The current plan dictates that each of us plan and cook two non-frozen pizza meals per week. The remaining three meals are "free." If we can keep this up for a reasonable amount of time, then we move onto baby step 2.

baby step 2: make the meals moderately healthy.

baby step 3: start brining lunch to work...maybe left overs.

The plan seems simple enough, but that doesn't mean we won't fail miserably at it. The continued commitment to exercise is promising, but seriously, we suck at this.

Brimstone and Fire

It is official. I am completely burned out.
I have lost all motivation to finish my project, write my papers or thesis, write in my lab notebook, make freezer stocks or use one more kit of any kind or sort. I do not want to extract, digest, ligate, transform, inoculate or incubate anything.
This downward spiral into the anti-motivation pit all started when I landed a post-doc. position. Just the thought of a new project, in a new lab at a different institution was all it took.
It started innocently enough. I put off those irritating tasks that must be done, but require very little brain activity to accomplish, like colony PCR or DNA extraction. Then I started doing half experiments. For example, I completed a growth curve (in triplicate) and froze away cells from various time points for later analysis and then I just left them in the -80. The samples are fine, but my point is that they are still in the freezer after three weeks.
The situation was only made worse by the fact that my boss marked off three weeks of vacation and out of town trips for the month of August (formerly known as the month I defend my thesis). So now, September is the month. No big deal. I am not upset or anything, but now the fire under my ass that I so desperately needed won't be lit for another couple of weeks.

The last couple of days have gone like this:
I wake up, I feel relatively motivated to go to work. While brushing my teeth I think to myself: I WILL do items X, Y and Z! Then I get to work and sit down at my desk and I revise the plan I formulated while brushing my teeth. Before I generate more information, I should really fill in my lab notebook so I don't get too far behind. BUT, before I can accomplish that, I decide that what I really, really need to do is analyze the data I currently have on hand. After all, it is data.

Rinse, repeat.

I know that I will return to my productive self and I hope it happens sooner rather than later. Maybe just admitting that I am a lazy bastard will help. We’ll see how tomorrow goes.