3rd June 2016 – Day 10

Friday:

Buffer day!! We made all of the buffers and solutions we are going to need for next week (in preparation for what we are planning to be a very busy week.)

1 litre of Buffer A – wash buffer

1 litre of Buffer B – elution buffer

One would think these buffers would be easy to make by following the instructions, but for some reason we all kept getting confused, it took us 3 attempts to get it right (at first we added the concentrations, instead of the volumes and then we put the opposite volumes of Imidazole in to each….really its like we were all having a momentary lapse of reason......they are only buffers). These were both pH’d to pH 7.6 using concentrated HCL. The pH meter had to be calibrated beforehand between pH 10 and 7. These will be needed for the pull down assays next week.

2 litres of gel filtration buffer

1 litre H20

1 litre 20% ethanol (made from 100%)

These last 3 solutions were filtered and degassed, it took a while but it needed to be done for the gel filtration column, it can’t have any bubbles (although who doesn’t love bubbles).

The column is stored in ethanol to stop bacterial growth, so it is washed with water and gel filtration buffer (this is to prepare it for use next week).

It's been such a busy two weeks, but I have really enjoyed every minute. My poor feet are looking forward to some downtime or should I say ''up time'' and a bit of rest this weekend, but I am really excited to see what next week holds.