Team:University of Ottawa/17 July 2008

From 2008.igem.org

(Difference between revisions)
Line 14: Line 14:
::<li> A transformation was performed on PTP2 ligation product - I then left the streaked plates to incubate for tomorrow.
::<li> A transformation was performed on PTP2 ligation product - I then left the streaked plates to incubate for tomorrow.
::<li> A glycerol stock was finally made of the BY4642 yeast strain int. DQ232597.
::<li> A glycerol stock was finally made of the BY4642 yeast strain int. DQ232597.
 +
'''Dan'''
 +
:'''Gel of construct 1 after ligation'''
 +
::<li> Gel was unsucessful, the wrong primers were used for PCR amplification.
 +
::<li> Ligation seemed to work very efficiently, digesting in 50 uL with the new ClaI enzyme is working very well.
 +
:'''Digestions'''
 +
::<li> Three digestions were performed 1+T, 1+T (higher concentration of vector and insert), and just the vector
 +
:'''Ligations'''
 +
::<li>Three above plasmids were ligated overnight

Revision as of 14:46, 22 July 2008

Contents

_

Today in the Lab

Chris

Purification of AtCRE
  • used PCR clean up kit to purify AtCRE sample
  • Tranformation of Competent Cells
  • followed transformation of competent cells protocol to insert AtCRE plasmid into preprepared competent E. coli cells.
  • allowed cells to incubate overnight at 37 C
  • Matt

    Ligation
  • As suggested by Dan the ligation was spiked with 1 ul ATP.
  • I then performed a PCR cleanup of the Ligation product in order to purify it for transformation into competent cells.
  • Transformation
  • A transformation was performed on PTP2 ligation product - I then left the streaked plates to incubate for tomorrow.
  • A glycerol stock was finally made of the BY4642 yeast strain int. DQ232597.
  • Dan

    Gel of construct 1 after ligation
  • Gel was unsucessful, the wrong primers were used for PCR amplification.
  • Ligation seemed to work very efficiently, digesting in 50 uL with the new ClaI enzyme is working very well.
  • Digestions
  • Three digestions were performed 1+T, 1+T (higher concentration of vector and insert), and just the vector
  • Ligations
  • Three above plasmids were ligated overnight