Friday, 4 July 2014

End of week 4

So its the end of my third week here in the Stockley lab, and its been a busy one!
Alongside moving house, I've been trying to complete my protease digestion experiment (mentioned in a previous post), which has proven to be more difficult then I first expected.

The protocol I am following is pretty straight forward, but with my first trail I encountered some problems. All my samples looked the same when analysed by SDS-PAGE, which meant possibly all my TCV sample was being digested by chymotrypsin as soon as I mixed the sample with the enzyme.  This is not what I expected or wanted to see!

Time to troubleshoot!!

First I tried to lower the amount of enzyme I added to the assay. No success.
Then I thought maybe my protein has degraded and that's why I'm not getting a result? So I tested for that on a gel alongside chymotrypsin on its own.
Conclusion? I think my protein is of very low concentration and can't by visualised by now I have to figure out where to go from here, especially as the amount of sample I have left is very little!

So this week hasn't had any major discoveries of science unfortunately, but that's okay because sometimes it takes a while for an assay to work! Also if I do a PhD, this is good practise for me, getting used to the fact that not everything in the lab will go right first time and sometimes you have to work on the same experiment for a while before getting a result! All experiences of what a real career in the lab will be like for the future!

No comments:

Post a Comment