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Team:University of Ottawa/18 June 2008
From 2008.igem.org
Contents |
Today in the lab
Dan and Matt
- Performed digestion of Tet plasmids
- T123, D12, and S1 were all digested with NcoI and EcoRV, results appeared to be as expected however EcoRI should have been used instead of EcoRV in order to obtain a better resolution
- Innoculated BY4741
- BY4741 was innoculated in 5 mL of 2xYPD + 1/10 20% glucose. However the BY4741 strain was 2 months old.
Chris
- Too few colonies grew on incubated plates; linear AtCRE may not have ligated properly
- Comparison of Ligated and Unligated Samples
- To determine of recircularisation worked, the ligated and unligated samples of AtCRE were run on a 0.8% gel for about 50 minutes. A difference was expected between the two samples when visualised.
- Neither sample turned up on the gel; concentrations of both were too low. This prompted a re-examination of AtCRE concentrations by absorbance.
- The concentrations of both samples were incredibly low; ligation did not work.
- Starting Over
- Digested AtCRE: 37 uL DNA, 1 uL EagI, 5 uL Buffer 3, 7 uL water.
