IGEM:IMPERIAL/2007/Notebook/Lab Planning

This page acts as a draft for the aims and projected achievements of the web lab... equipment requests and other information might also be present on the page.

Aims

Week 6:

• Clone remaining bricks in XL1-Blue
  • BBa_E0240 [GFP]
  • BBa_I13507 [RFP]
  • BBa_T9002 [ptet-luxR-pLux-GFP]
  • BBa_B0034 [RBS]
• Clone test contructs into BL21(DE3)
  • BBa_I13522 [ptet-GFP]
  • BBa_E7104 [T7-GFP]
• Midiprep bricks required for ligation
  • BBa_I13522 [ptet-GFP]
  • BBa_R0040 [ptet]
  • BBa_I0500 [pBad]
  • BBa_F2620 [ptet-luxR-plux]
  • BBa_F1610 [luxI]
  • BBa_E7104 [T7-GFP]
  • BBa_S03550 [RBS-araC]
  • BBa_B0015 [Stop]
  • BBa_R0051 [pcI]
  • BBa_E0240 [GFP]
  • BBa_I13507 [RFP]
  • BBa_T9002 [plux-GFP]
• Ligate constructs
  • Being planned
• Make cell extracts

Week 7:

• Clone bricks in XL1-Blue
  • BBa_J23039 [ptet-luxI]
  • BBa_I13504 [GFP]
• Clone test contructs into BL21(DE3)
  • BBa_B0015 [stop] (as a blank plasmid)
  • BBa_T9002 [plux-GFP]
• Ligate constructs
  • BBa_R0051 + BBa_I13504 = BBa_I719022 [pcI + GFP]

Outline of week:

• Monday: Preparation of pre-incubation solution and YT media
• Tuesday: Growing of BL21(DE3) in YT media
• Wednesday: Harvesting of BL21(DE3)
• Thursday: Preparation of cell extracts
• Friday: Preparation of cell extracts

Cloning-ligation cycle:

• Monday: Cloning of plasmids, plating
• Tuesday: Growing up of colonies for mini/midiprep
• Wednesday: Mini/midiprep
• Thursday: (Cloning, plating)
• Friday: Plasmid testing, (Ligation)

[Note: With an optimistic view, on cloning-ligation cycle takes 3 days.]

Eqiupment Waiting List

A list of media to be ordered and equipment to be autoclaved

Media

All media has been collected

Autoclave Equipment

• 10x 250 ml conical flasks
• 10x 50 ml centrifuge tubes