My F1: an update for all those curious blog friends

She arrived early (just a few days shy of the due date) on Thursday (1:30 in the am) after much ado about whether or not my water had broke. Silly me, I thought that would be something that would be pretty obvious, but it wasn't. By the time I got to labor and delivery (on Wednesday), I was informed that I had been in early labor for at least a day. Again, thought that would have been a little more obvious. As it turns out I am not that out of tune with my body, my water didn't really break, it just had a very slow leak and I have a fairly high threshold for pain.

Due to the slow leak, labor was induced and 13 hours later, I heard the words, "It's a girl." This coincided with extreme relief from pain and was promptly followed by the arrival of a slippery little person on top of my chest. My husband was great during the entire process and despite all the claims that there was no way he was looking "down there," curiosity got the best of him and he actually witnessed the birth. He declined to cut the cord though. I did not look. I figured, like with anything else that is painful, watching would only make it worse, so I kept my eyes shut and tried to focus.

We arrived home on Saturday and we've been adjusting to the new life ever since. So far, things are going well. The baby is still alive and intact and we haven't made any terribly huge blunders yet.

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 3 or Who am I kidding?

Don't get me wrong. I really am making the effort to be productive. I put in extra hours to make up for the trillions of doctors appointments I must attend and I try to keep my schedule at work so that I don't waste too much time while I am here. You know, the whole work smarter concept.

The problem is, I'm not smarter. I feel like my brain is on strike and I make a lot of really silly mistakes that I normally do not make. Let's take yesterday for example:

1. While performing a plasmid prep, I dumped all my samples into the waste container instead of onto the filter (which is where they needed to go).
2. I forgot to heat shock half my transformations. I went straight from ice to the plate.
3. I inoculated all of my overnight cultures into media containing antibiotics. Unfortunately, none of my strains were resistant to that antibiotic. In fact, these strains aren't resistant to any antibiotics.

So, yeah. Now I am even more behind. The cloning is already going very slowly thanks to the fact that E. coli fucking hates the constructs I am making. I can't really control this, but I'm doing what I can to minimize the problem. However, the things I can control, like say, using the correct antibiotic, I keep fucking up as well, making a process that is already going way too slow, move at a glacial pace.

Lab coat fail

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

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.