A day in the life of a PhD student. The Final Year edition.

I have kissed goodbye to those long days in the lab as a PhD student, which at the moment I am SO grateful about!

Why?

Well, I’m not gunna lie but those last few weeks, months perhaps, were tough! I worked my ass off to finish everything up in time for me to leave the lab and start writing my thesis which left me exhausted mentally, physically and emotionally. One of the aims of my blog are to share an honest account of my PhD experience so others might be more prepared for their journey or they have someone to reach out to when they might need support in similar situations, so I wanted to revive my ‘Day in the Life of a PhD student’ blog feature which I haven’t added to in a while and share one of my lab days in the last few weeks of my PhD lab time.

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This is not a normal day at all. I don’t want people to be thinking this is what needs to be achieved everyday. This day was from the last few weeks of my lab time in PhD and trying to wrap everything up. It is not healthy but I also want to share that sometimes it can get crazy busy if you are on a time restriction. Take a look back at other ‘day in the life of posts’ if you fancy it too πŸ™‚ So, let’s share with you a day in the life of a final year PhD student!

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7am: If you are a regular reader of Soph talks science and have read my ‘day in the life of’ posts before you will know that mornings and I don’t get on. Waking up is always a chore for me because I just love sleep but finishing up in the lab with experiments means lots of long days. And although I am 1,000,000% more productive in the afternoon evening I don’t really want to be in the lab all night so I’ve been forcing myself to be awake earlier than I normally would to start getting on with my day.

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8am: Up, dressed and samples for this morning on ice!

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8.40am: Would it really be a Soph talks science ‘day in the life of’ post if there wasn’t a Western blot? I don’t think so. So, yes I am still doing Western blots and I’m sure by this point you guys are as much of an expert in them as I am, but you can read up more about them here. Gels are made and samples prepped and loaded. Western blot is running and separating the proteins in my samples by size.

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8.50am: Samples thawed and ready for the next step of my chromatin immunoprecipitation assays which I am hoping will tell me if a particular protein binds to a region of the gene I’m interested in. In my ice box I’ve got two homogenisers like this one which are basically like a pestle and mortar. Time to put that elbow grease to good use and start lysing or breaking open my cells to release the DNA inside.

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9.45am: As if starting 6 new Western blots wasn’t enough for me today, I started 4 yesterday and need to develop today! They don’t call me the Western blotting queen for nothing haha. I think I have too many samples to run and too many antibodies to test them with hence the Western blot juggling but it means more exciting results for me – hopefully!

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10am: Setting another gel now. Not for another Western blot you will also be glad to know, but instead to separate DNA products on and test whether some cDNA samples I prepared yesterday were contaminated or not.

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10.30am: In the time the gel took to set, I finally managed to log on and check some emails, but now its ready, time to load my samples and hit ‘RUN!’

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10.45am: Time to wash yesterday’s Western blots and head to my second home – the dark room – to develop them.

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11.15am – While those blots are developing in the dark room, time to run back to the lab and take care of today’s Western blots. Samples are separated and now time to transfer them out of the gel and onto a membrane.

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11.45am: Back to the dark room to find out if my Westerns had worked, and its good news! Beautiful protein bands! Now to look for actin expression so I can quantify them.

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12.10pm: After a long and slow run, all my samples have reached the bottom of the gel and separated out again by size. But I can’t see just by looking at the gel here if my samples were contaminated or not, so time to light it up and see.

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12.15pm: So when I made my gel, I added a compound to it that binds to DNA and when I shine UV light onto the gel will illuminate where my DNA bands are.

A quick snap and this is what I see! The lane on the far left shows my marker where each band represents a particular size of DNA fragment and helps me to work out the size of the DNA fragments in my samples. So, most of my PCR reactions worked but there is no contamination in any of them. Yay! All the bands are of a particular band size. If there was contamination, there would be a DNA fragment of a a larger size and so would be higher up on the gel compared to these. But these samples are not contaminated and ready to be used in PCR reactions now!

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12.55pm: More samples on ice to thaw ready for this afternoons experiments while I take a while to sit and refuel.

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2.45pm: Lunch – eaten. Todays Western blots transferred and now into block. DNA from my chromatin immunoprecipitation experiments sheared. And the first of todays PCR plates ready to be run for analysis.

Plate loaded. Layout set. Run started. Now back to the lab.

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3.20pm: Okay, this isn’t deja vu. But I’ve set another gel but this time to check that my DNA from my chromatin immunoprecipitation assay has been chopped up enough to move on to the next steps. Samples loaded and time to hit run again.

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3.40pm: Time to wash and develop yesterdays Western blots again for actin expression so I can analyse the expression of my other proteins of interest in my samples. You guys definitely know the drill here – wash, develop, head over to the dark room, hope that protein bands appear.

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4.10pm: Got protein bands! Woo! And DNA gel is still running so time for another lab daily ritual – stem cell culture! Still have quite a few plates of stem cells to look after and use in experiments, so this means a lot of sample collection and setting up experiments whilst also singing to my cells to make sure they stay happy. Genuinely. πŸ˜› I’m convinced it helps!

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4.40pm: Time for a quick switch around and a ‘here’s one I prepared earlier’ moment! PCR plate 1 is finished so taken the data to analyse later and run plate number 2 of the day.

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4.55pm: DNA gel number 2 complete and time to head down to check the DNA bands out again.

These bands are different to the gel earlier in the day because rather than a single band for each sample, I want to see multiple bands for each one. Why? Because this time around I want to see that my DNA is sheared enough and in the shearing process the DNA gets chopped up randomly and so we expect to see an array of different DNA lengths, rather than the one positive or negative band when checking for contamination earlier. The sheared DNA all needs to be within a certain range of size and although its not particularly easy to see in this photo, both samples have DNA that has been cut up enough for me to use in my immunoprecipitation experiments. The downside is there isn’t much DNA in my samples.

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5.45pm: A quick clean up of those sheared DNA samples and time to set them up for my immunoprecipitation experiments. I’ve added magnetic beads to my DNA and added an antibody that binds to my protein of interest and allow them to all mix together on this rotator overnight. If my protein does bind to the region of the gene I am interested in, then it will be holding onto that piece of DNA in these samples. Then in the next steps of this experiment, I can pull that piece of DNA out of the mix and then run another PCR to determine if it is that gene region.

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6.10pm: Time to wrap up todays Western blots too. Membranes added to these tubes with yet more antibodies to analyse the expression of proteins that I am interested in in these samples. Then to leave them chilling and rolling overnight too ready to continue tomorrow which you know what is involved with now too πŸ™‚

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6.45pm: After analysing todays data and writing it all up in my lab book, it’s time to collect the data from PCR number 2 now that’s finished and get all that analysed too.

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8.15pm: All lab work for today written up in my lab book. PCR analysis done. To Do list for tomorrow done! Equipment booked. Time to finally head home and refresh ready to do it all again tomorrow!

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As I said at the start, these final few months of my PhD were tough. I wanted to do everything I possibly could to finish that thesis story that I am now writing about but it definitely was draining. And if you are in the final year of your PhD and thinking ‘I’m not doing this much’ please do NOT think that you have to. Instead do whatever is best for you and your research. Each PhD and PhD student is different so don’t feel like you have to fit in.

But no matter how much I love being in the lab, I am so glad that these days are over and I can start to think about getting healthier, thinking about that post-PhD life and even start to get excited about planning a wedding!

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If you want to know more about any of the experiments I was doing on this particular day then please ask questions about them below or contact me on social media. Or perhaps you want to share a typical day in the lab for you to help me show that no two PhDs are the same and you need to work to whatever YOU can or want to do. You can either do that in the comments, write your own blog or maybe even write a guest post for me πŸ™‚Β What is a typical day like for you and your science research?Β Please share your stories so anyone who reads this blog can see how varied a scientist’s day can be! It is not lab 9am-5pm Monday to Friday.

Science love.

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