Afternoon All,
Quite a quick day today! Myself and Mike viewed the PFGE gel we ran last Thursday to Friday. Although the run appeared better in terms of visible band separations, the bands were still fairly faint and the wells appeared very dark. Mike said that this was due to too much DNA being left in the wells, we therefore decided to reduce the amount of plug we put into the well, by cutting the plugs as small as we could. We decided to reduce the buffer %, from 1% to 0.5%, to get rid of the initial 3 hour with 15 second switch time at 2.0 V/cm and add a second band marker (1Kb ladder). This 1Kb ladder was added as a reference in order to help us work out how many bases were present within samples.
We set up a PFGE run with all these new changes for 24 hours at 2.0 V/cm with a 45 second switch interval, and a second block for 1 hour at 0.6 V/cm with a 45 second switch interval in case we weren’t ready tomorrow morning to remove the gel. So we will see tomorrow what our changes have done! 🙂
Mike also showed me the TGYE agar he was making up, in order to re culture the Propionibacterium acnes samples. He showed me how to make up the agar, autoclave it and pour it. Tomorrow he is going to show me how to streak plate the Propionibacterium acnes samples onto the TGYE agar plates in the anaerobic cabinet. I have never used it before, so it should be really good to see!
See you all tomorrow! 🙂