Imperial/Wet Lab/Lab Notebook/2007-08-14

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__NOTOC__
= 14 August 2007 =
= 14 August 2007 =
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==Preparation of Stock Solutions==
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*1x 1 litre of 2xYT medium in Duran bottle
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*3x 1 litre of 2xYT medium in conical flask
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**Sent to autoclave
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*1x 10 ml of pre-incubation solution for preparation of cell extract
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*6x 20 ml of 1mM IPTG stock
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==Preparation of Cell Extract==
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#Picked a BL21(DE3) colony from Tet plate
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#Innoculated in 100 ml of 2xYT + Tet medium
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#Incubated overnight at 37 °C
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==Cloning of Biobricks==
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#Placed 5 ml of LB in a 15 ml tube
 +
#Added appropriate antibiotics into the tube
 +
#Picked a colony from the fresh overnight plate
 +
#Innoculated the colony in the LB media
 +
#Incubated for 6 hours during the day
 +
#*2x 6 Biobricks cloned
 +
 +
==Cloning of Biobricks (for Midiprep)==
 +
#Placed 50 ml of LB in a 250 ml conical flask
 +
#Innoculated 1 ml of culture into LB media in flask
 +
#Incubated overnight at 37 °C
 +
 +
<font size=-2>
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*2. BBa_I13422 [ptet-GFP]
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*5. BBa_R0040 [ptet]
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*7. BBa_F2620 [plux]
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*8. BBa_F1610 [luxI]
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*11. BBa_E7104 [pT7]
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*13. BBa_B0015 [stop]
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*14. BBa_R0051 [pcI]
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*16. BBa_E0240 [GFP]
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*17. BBa_I13507 [RFP]
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*18. BBa_T9002 [plux-GFP]
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==Pilot Preparation of Vesicles==
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*The suspension prepared the [[Imperial/Wet Lab/Lab Notebook/2007-08-13 | day before]] was used to produce vesicles enclosing Tris and NaCl buffer only.
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*There was a mistake in following the protocol: instead of preparing the interface with 2ml of the DOPC-mineral oil suspension and then adding 100&mu;l of emulsion to it, the emulsion was prepared according to protocol, and then 2ml of emulsion was used to create the interface.
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*'''Results''': A sample was briefly looked at under a light microscope, but no vesicles were observed.
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Latest revision as of 12:42, 26 October 2007

14 August 2007

Preparation of Stock Solutions

  • 1x 1 litre of 2xYT medium in Duran bottle
  • 3x 1 litre of 2xYT medium in conical flask
    • Sent to autoclave
  • 1x 10 ml of pre-incubation solution for preparation of cell extract
  • 6x 20 ml of 1mM IPTG stock

Preparation of Cell Extract

  1. Picked a BL21(DE3) colony from Tet plate
  2. Innoculated in 100 ml of 2xYT + Tet medium
  3. Incubated overnight at 37 °C

Cloning of Biobricks

  1. Placed 5 ml of LB in a 15 ml tube
  2. Added appropriate antibiotics into the tube
  3. Picked a colony from the fresh overnight plate
  4. Innoculated the colony in the LB media
  5. Incubated for 6 hours during the day
    • 2x 6 Biobricks cloned

Cloning of Biobricks (for Midiprep)

  1. Placed 50 ml of LB in a 250 ml conical flask
  2. Innoculated 1 ml of culture into LB media in flask
  3. Incubated overnight at 37 °C

  • 2. BBa_I13422 [ptet-GFP]
  • 5. BBa_R0040 [ptet]
  • 7. BBa_F2620 [plux]
  • 8. BBa_F1610 [luxI]
  • 11. BBa_E7104 [pT7]
  • 13. BBa_B0015 [stop]
  • 14. BBa_R0051 [pcI]
  • 16. BBa_E0240 [GFP]
  • 17. BBa_I13507 [RFP]
  • 18. BBa_T9002 [plux-GFP]

Pilot Preparation of Vesicles

  • The suspension prepared the day before was used to produce vesicles enclosing Tris and NaCl buffer only.
  • There was a mistake in following the protocol: instead of preparing the interface with 2ml of the DOPC-mineral oil suspension and then adding 100μl of emulsion to it, the emulsion was prepared according to protocol, and then 2ml of emulsion was used to create the interface.
  • Results: A sample was briefly looked at under a light microscope, but no vesicles were observed.


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