27 and a PhD

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150 mM NaCl and the PMDD acting up

Welcome to my blog!

Hello there, awesome reader. My name is Dr. 27. I'm older than that now, but I'm staying faithful to the origins of the blog.

This blog started 2 months before completing my PhD in a pretty southern university back in 2009. It was a way to practice my writing and take a break from all things thesis. My PhD is in a branch of structural biology where I studied some rather impressive stuff.

After completing the degree, I packed my life of 6 years in 3 days and moved to Canada to do a postdoc in a completely different field. Two years later, and after attending a lot of seminars, workshops and doing some much-needed soul-searching, I ended up getting out and looking for an alternative path to academia and industry.

The blog chronicles my mishaps, ideas, musings and tips on entering, staying and finishing grad school. It also talks about some (or a lot) of personal stuff. For a while, the blog became a place to talk about the frustrations of not knowing what to do after PhD. I wanted to explore alternatives to the traditional paths of research (academia, industry and goverment) whilst going back to my field of training (if at all possible). Eventually a job materialized. Follow my quest as I navigate the waters of being a staff scientist at a core facility.

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I know I’ve been absent from the blogging scene for a little while. So many things have happened in the last 3 weeks it’s been crazy. And anytime I get a chance to log in, approve comments and catch up on little things I end up dreading writing a post about current events.

Part of the reason I’ve been out of the loop is due to a recent discovery. Since January I’ve been trying to purify this protein which is one of the main building blocks of my postdoctoral research. Said protein was made as a fusion protein and I was given a protocol to follow which is similar to others used by the group. My lab is very successful at expression protein (same as many others) and the tools to grow, purify and obtain mg amounts of protein are pretty standardized.

Enter moi. I was a bit …. mmm, spoiled while in grad school. As I didn’t have to do much biochem to obtain my samples of interest (sometimes people would send their so our facilities could analyze them), I forgot all about purifying, growing and all those other details which make biochem a necessary discipline of the trade. Thus, I “warned” my new boss of my biochemical incapacity and he agreed I’d have a few months to try and purify things on my own and do it proficiently.

As I mentioned in paragraph #1 my lab is very successful at purifying proteins so I felt I had little to fear. Now, I’m a very careful person, but if someone else has designed (and tried on a very small-scale) a purification protocol, based on experience I’ll follow it to a tee and usually it works.

Enter March 2010. All of my biochem frustrations and fears and you name it came crawling and chasing me as column, after column, after column, all I got was my desired protein PLUS the cut tag. The friggin’ thing would not, could not come off.

Earlier this week I had a fall out with Dear Boyfriend in which I told him all I thought … about me, and how incapable I was feeling. BUT earlier that day I decided to try adding 150 mM salt to my buffer and see how it went the next day.

Ba da min! Ba da boom! IT WORKED! I texted Dear Boyfriend to communicate the happy news (he had seen how frustrated and mad I was, yet remained as encouraging as always). I sent the protein for more analyses and it’s not degraded (yee pee) and now I think I have it all worked out.

On the scientific side things are looking promising.

On the personal, or mental state side things aren’t as bright. PMDD is acting up … and badly. My mood has been in the trash for almost a week, even though my lab stuff is working. I’ve been snapping at the boyfriend and any little provocation and I feel like an ogre.

SO, between troubleshooting and buying the PMDD med and having it take effect my blogging capacity is little to none existent. I’m feeling miserable dear blogging world, and for that reason I’ve been neglecting it. Please don’t take it personal, it’s all me and my head.

Soon I hope to be back in full speed and have lab and life things under control. Till then, dear interwebz don’t think I’m depriving you of my talent. It’s a temporary thing which I hope to solve or at least gain some control on very soon.

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3 Comments

  1. Alexandra says:

    Congratulations for getting it to work. Science can be so weird, eh? Sometimes you just have to pipette with your left hand and it will work 😉

  2. 28 says:

    Thanks a bunch! Now the hard part is forming the complex I need. Thanks for visiting 🙂

  3. […] – Had a problem with one of the purification columns  but eventually it was fixed. Figured out that 150 mM salt was needed for my favourite tagged protein to bind to another column (I learned about how important it is to […]

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