IGEM:Paris Bettencourt 2012/Notebooks/suicide group/day by day//2012/08/06: Difference between revisions

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overnight (saturated)
overnight (saturated)
*Dilute 100x; plate 20uL with beads
*Dilute 100x; plate 20uL with beads
*On each plate put drops of saturated WT, ColE2 and ColE7
*On each plate put drops (5 uL) of saturated WT, ColE2 and ColE7
 
===Result===
===Result===
ColE2 seems stronger to ColE7; WT more immune to ColE7
ColE2 seems stronger to ColE7; WT more immune to ColE7

Revision as of 02:33, 7 August 2012

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Gel purification

(because I did the digestion few days ago, so we need to check if it does not cut non specifically)

  • pSG.004 (X,P)
  • pSG.014 (X,P)
  • pSG.020 (X,P)

Ligation

Product

Product Description Vector Insert Antibiotic resistance
PSG.003 (-) Negative control (vector only) PSG.003 (S, P) - Amp
PSG.021 pSB1A2 pLac ColE2Imm PSG.003 (S, P) PSG.014 (X, P) Amp
PSG.022 pSB1A2 pLac ColE7Imm PSG.003 (S, P) PSG.020 (X, P) Amp

Protocol

Vector (32 ng) 2 uL
Insert (20 ng) 5 uL
Buffer 2 uL
Ligase 1 uL
H2O 10 uL
  • Incubate 1h in the microscope room (20°C)

Transformation

only 1 colony, on the (-) control plate :(

Colicin experiment

Protocol

  • Grow cells
    • Wild type
    • ColE2
    • ColE7

overnight (saturated)

  • Dilute 100x; plate 20uL with beads
  • On each plate put drops (5 uL) of saturated WT, ColE2 and ColE7

Result

ColE2 seems stronger to ColE7; WT more immune to ColE7

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