Today was much busier than yesterday, there was a lot to get
done and I learnt a lot of new things which was exciting.
The overnight cultures were put through an Axygen-Axyprep
Plasmid miniprep spin (kit). It turned out to be a frustrating start to the day
as the initial prep kit we wanted to use was empty and so we wasted time fishing
around to find a different one.
I prepared larger cultures taken from falcon tubes into
conical flasks, these were incubated and the optical density was measured (OD -
to find out how many cells are actually floating about, we want them in their
log phase where it will be most efficient. Between 0.4 and 0.8). Once required
OD was reached, I induced them with IPTG and incubated overnight. I took
samples before and after inducing so it could be run on an SDS-Page gel.
Everything I have done up until now I had only learnt about
in theory so actually doing it was exciting but at the same time a little daunting.
I have run an SDS-Page gel before now, so this at least was not completely new,
however before I could actually run the gel, I had to make it……from scratch (normally
the technicians prepare these before class, it’s a bit like being on Blue Peter
when one magically appears and they say ‘this is one we made earlier’). I did the resolving first and let it set
before I prepared the stacking gel, there are a lot of steps to go through and
if one thing goes wrong, it has to be started
again (its not so scary once you get started).
Once this was all completed and the gels were placed in the
fridge to store over night, I transformed my cells and these were left to
incubate for an hour. I then plated them and left them to incubate overnight (I
have done a streak plate before now, but not this many all at once).
Well, day 2 done, there was a lot more waiting around today in
between the incubation times which was nice as I got a chance to sit down (but definitely
a good call on the trainers…phew!).
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