IGEM:IMPERIAL/2006/LabCalendar/2006-9-12

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TO DO:
*Maxiprep & Minprep J37025 & J37022 ? (Depends on results of J37024 control miniprep from yesterday)
*Electroporate the overnight ligation of J37024 from Dr. Mann
*Sequencing 5M, Aiia, (022 & 025)
*Analyse the Western Blot for protein (Aiia) expression
*Biosensor testing (Biosensor Calibration)
*Culture J37016 for contd. Biosensor testing (Enzyme Activity)

Contents

Sequencing

aiiA-B and the maxiprep of aiiA from the registary were sent off for sequencing

Western blot

This didn't work. There is no diference between the controll and the test (both circled in white). However the strain used can only express one of the promoters and there was background expression reported by Dr fray, both of these could hide a positive result.

Finish ligation of J37024

  • 10μL of the J37024 ligation was left to ligate overnight
  • Glass Milk DNA purification of this was carried out
  • Electroporation - check tonight whether we already have any colonies on the plate

Biosensor Testing

  • The biosensor was tested according to the Biosensor Calibration Testing Protocol
  • Rows D, D Control, F, F Control, G, G Control, Control LB, Control Enzyme were tested
  • Both LB and MiliQ were tested as medium
  • RESULTS:
    • All pH readings in LB were in the pH range of 6.31 and 6.35 - with the control and the actual test (where the enzyme was added) being mostly the same value
    • pH readings in MiliQ were in the pH range of 5.90 - 6.53. However, no significant difference could be detected in the readings of the test (where enzyme was added) and the respective control
  • NOTE: The initial measurement of MiliQ was pH 7. However, after measuring the samples, it was at pH 6.70. Next time, calibration in between the readings should be made.
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