IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010/2010/09/05

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Aleksandra
Touchdown PCR of the 13 minipreps from yesterday and also of 6 other constructs (elongation time 6 mins), then gel electrophoresis.


Gel 1 (PCR)
Minipreps from 31/08

  • 1 Pms/wTet II
  • 2 Pms/mTet III
  • 3 Psm/mTet III
  • 4 Psw/mTet III
  • 5 Pss/wTet II
  • 6 Psw/wTet II

Minipreps from yesterday (04/05)

  • 7,8 Pms/mTet/LuxI, 2 cultures
  • 9,10 Pss/wTet/LuxI, 2 cultures
  • 11,12 Psm/sTet/LuxI, 2 cultures
  • 13,14 Psm/wTetIII/I13507/pLux/GFP, 2 cultures
  • 15,16 Psm/wTetII/I13507/pLux/GFP, 2 cultures
  • Ladder 1 kb

File:PCR0509.jpg


Gel 2 (PCR)
Minipreps from yesterday (04/05)

  • 17, 18 Pss/sLac/LuxI III, 2 cultures
  • 19 Pms/sTet/LuxI

Digestion from today(see further)

  • 19 Pms/sTet/LuxI

File:PCR0509 2.jpg


→ Colonies 7, 9, 10, 17, 18, 19 seem to be of a right size


Restriction digest and gel electrophoresis of yesterday's minipreps with E+P
Gel 1

  • 1 attP/PmsRFP, col 1
  • Ladder 1kb
  • 2 attP/PmsRFP, col 2
  • -
  • 3 attP/PsmRFP, col 1
  • -
  • 4 attP/PsmRFP, col 2
  • -
  • 5 attP/PssRFP, col 1
  • -
  • 6 attP/PssRFP, col 2
  • -
  • 7 Pms/mTet/LuxI, col 1
  • -
  • 8 Pms/mTet/LuxI, col 2
  • -
  • 9 Pss/wTet/LuxI, col 1

File:Gel0509 2uncut.jpg


Gel 2

  • 10 Pss/wTet/LuxI, col 2
  • -
  • 11 Psm/sTet/LuxI, col 1
  • -
  • 12 Psm/sTet/LuxI, col 2
  • -
  • 13 Psm/wTetIII/I13507/pLux/GFP, col 1
  • -
  • 14 Psm/wTetIII/I13507/pLux/GFP, col 1
  • -
  • 15 Psm/wTetII/I13507/pLux/GFP, col 1
  • -
  • 16 Psm/wTetII/I13507/pLux/GFP, col 2
  • -
  • 17 Pss/sLac/LuxI III, col 1
  • -
  • 18 Pss/sLac/LuxI III, col 2


File:Gel0509 1uncut.jpg
File:Gel0509 1cut.jpg

  • 19 Pms/sTet/LuxI (on the PCR gel, see above)


→ Digestions 1, 2, 7, 9, 10, 17, 18, 19 seem to be of a right size