Wednesday, March 3, 2010

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.


PhizzleDizzle said...

you are a machine. i think if i were pregnant i would only plan to make mr. phizz go get me food and not think much about anything else ;).

Dimitris said...

i bow to you. really.

ScienceGirl said...

Nothing like the motivation of the final stretch! Good luck with all that (although don't be surprised if you get progressively more tired - I know I am beyond the point of possibly being productive).

Thomas Joseph said...

26 strains for one paper? Sounds like you could really get two manuscripts out of such a study.

microbiologist xx said...

PD - I try to be a machine at work, but at home, I am pretty much as you imagine yourself to be. There is a lot of laying on the couch. :)

Dimitris - Aw shucks. Let's just see if I can get this stuff accomplished in the allotted timeframe. If I can, then by all means bow away. ;)

SG - You are absolutely right. I am feeling very motivated. I am really going to give it the ole' college try, but not at the expense of my health or the baby's health. The fatigue is starting to set in already, and I don't see myself doing much of anything at all if I am past my due date.

TJ - Yes. It sounds crazy, but it is true. Those 26 strains represent two sets of 13 strains (two different backgrounds). There is every possibility that I won't need all 13 of the second set, but I don't have the luxury of waiting and seeing how experiments with the first set go before I start making set two, so I just decided to bite the bullet and make them all, knowing full well that I may not need them all. Besides, it's only cloning, right? On the plus side, some of these will be used for future studies (and papers) too.