#PhDaily Day 2 11/08/2015

Hi everyone,

A slightly earlier start this morning of 9.30, after a shorter but seemingly better night’s sleep! Shortly after submitting last night’s post, a fuse tripped in my house and wouldn’t come back on. After eventually fixing it (somehow!), it was quite late, so I had a late night.

The plan this morning was to start an experiment between 9 and 10 am, as i had a lab meeting to attend at 10. However i needed several of my supervisors’ stocks, and as he wasn’t in, i couldn’t access them before the meeting! The same undergrad i set up the FPLC for last night was ready to inject one of his samples, but another supervisor in the lab noticed how the equilibration step hadn’t worked properly. He explained that i hadn’t set up the equilibration for long enough, and just needed to modify my protocol in the future! Thankfully this was a quick fix, we ran some more buffer through the column and it was good to go. It is helpful when things go slightly wrong, as this is when i usually learn the most!

It was a few minutes until the lab meeting at 10 am, so i ran down the three flights of stairs and across a courtyard to my other lab. As i do protein NMR, i am co-supervised, so have another lab too! This also means i have to (try to) attend two lab meetings each week. Today was my turn to present to the NMR group. I gave a ‘short’ presentation – mainly focusing on things that haven’t worked – which after conversation and questions, ended up taking almost an hour!

They are a gracious but small audience, applauding me twice – once when i tell them about winning best student talk at the SfAM summer conference, the other when i finished my talk! I stay after finishing for a quick chat with some friends, then make my way back upstairs to the microbiology lab. I interrupt my supervisor (who is dealing with the FPLC again!) to find out where the stocks are that i need.

The reason i wanted to start the experiment so early this morning was because i am enzymatically digesting peptidoglycan (the cell wall of bacteria), for an important experiment next week. The type of experiment that could make a figure for a paper… These digests take approximately 6 hours, at which point the enzymes are topped up, before leaving overnight. My supervisor, noticing the time, ensures me that i can just add the enzymes before going home.

I filter the buffers i need for this process, a: because it is quicker than autoclaving, and b: because the autoclaves are currently in short supply in our building! I then set up the digests, just some simple pipetting following a recipe, followed by incubating them at 37 C. I want to go on my lunch, but the undergrad is ready to do their second injection of the day on the FPLC. I help them out, managing to leave for lunch by just before 2pm.

I walked down the road to get a panini from one of my favourite shops, then eat back in the office while catching up on news again. I also tend to my various twitter accounts, and celebrate because my personal account reached over 1000 followers today! At about 3pm, my supervisor comes to the office for a quick chat to plan my important experiment next week. I answered a few emails (it’s good to keep on top of them), and then answer a few more questions one of the undergrads had. I didn’t get too involved with the last set of undergrad students we hosted in the lab, as it was back in October-December when i had just started my PhD. Now i am a bit more confident, and i actually have some of the knowledge they need! I take the rare opportunity of things being quiet in the lab to write up some of this post, and also work on an article for the Sheffield University Student’s blog, to which i am a regular contributor.

At around 4pm, i head back into the lab to do some tidying up, make a few more buffers and check my stocks. For my final supervisory duties of the day, i helped set up the cleaning program for the FPLC. In a nice way to end the day, i spent a bit of time just chatting and joking around with a couple of the undergrads and another PhD. I added the second batches of enzymes to my peptidoglycan digests at 6pm, then cycled home. Tonight I think i will wind down by playing some video games!

Thanks for reading,

Microbe Stew

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