31st May 2016 – Day 7

Tuesday:

Two new volunteers in the lab today, and Amy charged me with showing them the ropes and overseeing things. I know them both well from class so at least we are all familiar (but one thing I have learnt today is I definitely like being in charge!).

We didn’t do anything new today, it was all similar methods to last week, but it allowed me to build some confidence as I knew what I was doing and what the processes were. I showed both volunteers how to make SDS-PAGE gels, we made 4 and prepared the 10% APS on our own. 

We got a chance to photograph the gel that I ran on Friday (the one that had been de-staining over the weekend). This worked really well (but note to self don’t make gels symmetrical with the marker in the middle, there is no way to tell which way round it should be, d’oh!).

The gel dried out too much and it caused it to become brittle.

We made more buffers, ones we will be requiring for the western blot at a later date. We made a litre of Transfer buffer and a litre of fresh 10x TBS buffer, which we had to pH using a probe. I never realised how hard this would be, it took forever to get the buffer to the correct pH (7.6) to be exact. The probe was calibrated at the start and washed well between each solution.

We did a streak plate of SUFU using the glycerol stock, we will be using this to make up larger liquid cultures in the future. We made two small liquid cultures of 5ml LB each, 2 using KAN and these were incubated overnight. We prepared 2m 100ml liquid cultures in conical flasks and autoclaved them in preparation for tomorrow.

It was a very productive day, we got through loads of work and it definitely helps having the extra pairs of hands.