This week I have been learning how to do mini preps (plasmid purification) independently. The process includes lots of pippetting, centrifuging many buffers that serve different purposes, but in the end help to purify our plasmid product. After the plasmid is purified, the concentration of nucleic acid in the sample is measured using a Nanodrop machine. Once this is completed, the sample is ready to be sent off for sequencing. This sequencing allows us to identify into which sites the plasmid has incorporated our non canonical amino acid (ncAA) mutation. Once those sites are known and isolated, it allows us to plate plasmid containing bacterial colonies with a single site mutation at different temperatures. I must say, I love being in the lab and getting the privilege to play even a small part in this work because its broader implications could be massive, and the work is equally as fascinating as it is challenging.
The first photo below shows an apparatus that is hooked up to a vacuum where wash buffers are used for the plasmid purification. And the second picture shows the Nanodrop machine.
The nanodrop machine is cool! Is it the one in BRC? I remember having to be super extra careful and only needing a single drop. I remember doing mini prep too but in a different way. What’s the bigger picture for your project? I never got to do the tissue culture since our plasmid was not perfect when we sent it for sequencing.
I love doing Mini Preps!… and using the nano drop. We’ve been cruising along with the sequence of protocols to make our virus and haven’t done one since about week 1 =(