Sp'23: Week 13 (4/17/23 - 4/21/23): Preparation and Readings

This week was more focused on planning, but I was able to perform a few tasks to continue forward with my project. 

First, as it is good to have fresh LB and TGY to pull from, I went ahead and created two new microcentrifuge tubes (1 mL each) of TGY and LB broth to pull from for nanodrop readings when needed, with the following tube names: 

  • "TGY 4-21-23 M.O. 4.21A" 
  • "LB 4-21-23 M.O. 4.21B" 
With this, I wanted to check the growth of the two inoculations I did the Friday before. Each of these tubes held LB + Ampicillin [with an E. coli mic concentration of 50 ug/mL] and were inoculated with E. coli that held the pRad1 plasmid. Unfortunately, the OD600 readings were extremely low (negative numbers), which indicated either no growth or death of the E. coli that may have grown (as they had been in the 37 degree Celsius incubator for almost a week). The OD600 values were as such: 

Now, because the OD600 readings showed that there were likely no E. coli alive in either tube 4.14A or 4.14B, I went ahead and reinoculated E. coli with pRad1 from a different plate (plate 1.23D rather than 1.23C) into two new tubes with LB + Ampicillin [mic concentration of 50 ug/mL], labeled "4.21C" and "4.21D," with the hopes that this would result in growth of E. coli with the pRad1 plasmid in the two new tubes. 

These two tubes were then placed into the shaking incubator at 32 degrees Celsius for hopeful growth over the weekend: 

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