Sometimes science can feel really slow. I can be stuck waiting for things to happen, cells to grow and deliveries to come for weeks on end. Then sometimes, I have a week like the one just past.
I hatched a plan for a big study with Manos, my supervisor, before Christmas and, after several protocol changes and refinements, failed attempts and major rethinks, we’re finally getting going. I am very excited about this study – I was bouncing around the lab when I took delivery of seven Nucleospin kits by Macherey Nagel (via Fisher Scientific). If you’re interested in simultaneously extracting RNA and protein from your samples, the Nucleospin is for you. I love the extra validity of doing both extractions from the same cells and it’s a really simple and robust protocol to boot.
The plan is to extract conditioned media, RNA and protein from three cell lines treated with inhibitors, at two concentrations, over three time points and, of course, in triplicate. Manos did warn me that it would be a lot of work and that I may want to consider removing a time point or concentration but the lure of covering all bases was too much. I knew I’d have to push myself to the limit to get everything done but I wasn’t fully prepared for how the week would go.
Taking one cell line at a time, the idea was to seed into 6-well plates on the Monday, treat with six different inhibitors on the Tuesday and then do 24, 48 and 72 hour extractions on the Wednesday, Thursday and Friday respectively. The seeding was fine, the treating not too much of a problem but in the 72 hours that was Wednesday, Thursday and Friday I spent around 30 hours in the lab. Like I said, the protocol is simple and robust, but when you’re working with 36 samples at a time, even loading them into the centrifuge can become laborious. I used well over 1,000 tubes over the course of five days and blisters were forming on my thumbs from popping them open. I was tired, hungry, dehydrated and using pipette tips faster than they could be autoclaved but by the end of the week I had 216 conditioned media samples, 108 RNA samples and 108 protein samples in the freezer.
An achievement, I think you’ll agree.
I took full advantage of the bank holiday weekend, made it a full week and chilled out with some friends in London before getting back to it. Only two cell lines to go!
Karly first joined BJS Biotechnologies as an undergraduate placement student in 2007. A bachelor’s and master’s degree later and Karly is back working with BJS but this time as a PhD student in conjunction with her alma mater, Brunel University. Her research looks at the mTOR signalling pathway in endometriosis and ovarian cancer.
In her spare time, Karly is learning to play the guitar and drinks a lot of tea.
You can follow Karly on twitter @KarlyBroadway