Team:University of Ottawa/18 June 2008

From 2008.igem.org

(Difference between revisions)
(New page: __TOC__ ==Today in the lab== '''Dan and Matt''' :'''Performed digestion of Tet plasmids''' :<li> T123, D12, and S1 were all digested with NcoI and EcoRV, results appeared to be as expected...)
Line 6: Line 6:
:'''Innoculated BY4741'''
:'''Innoculated BY4741'''
:<li>BY4741 was innoculated in 5 mL of 2xYPD + 1/10 20% glucose. However the BY4741 strain was 2 months old.
:<li>BY4741 was innoculated in 5 mL of 2xYPD + 1/10 20% glucose. However the BY4741 strain was 2 months old.
 +
'''Chris'''
 +
:<li>Too few colonies grew on incubated plates; linear AtCRE may not have ligated properly
 +
:'''Comparison of Ligated and Unligated Samples
 +
:<li>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.
 +
:<li>Neither sample turned up on the gel; concentrations of both were too low. This prompted a re-examination of AtCRE concentrations by absorbance.
 +
:<li>The concentrations of both samples were incredibly low; ligation did not work.
 +
:'''Starting Over'''
 +
:<li>Digested AtCRE: 37 uL DNA, 1 uL EagI, 5 uL Buffer 3, 7 uL water.

Revision as of 15:14, 28 July 2008

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.
    • Retrieved from "http://2008.igem.org/Team:University_of_Ottawa/18_June_2008"