Team:Paris/August 1

From 2008.igem.org

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Revision as of 17:08, 13 August 2008

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Tomorrow →


Contents

MiniPreps

Conditions

  • Use of Quiagen's protocol on all the clones cultivated on the 31th.
  • Preparation of 50µl of minipreps in water.


List of Minipreps

Name Biobricks Description
MP124 Strong rbs - YFP
D129 (BV) - D118 (BI) 		Part icon rbs.pngPart icon reporter.png
MP125 Strong rbs - mRFP
D129 (BV) - D122 (BI) 		Part icon rbs.pngPart icon reporter.png
MP126 Strong rbs - lasR activator
D129 (BV) - D114 (BI) 		Part icon rbs.pngIcon coding.png
MP127 Strong promoter - ECFP
D123 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP128 Strong promoter - gfp Tripart
D123 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP129 Strongest promoter - ECFP
D103 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP130 Strong promoter - gfp Tripart
D103 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP131 Medium promoter - gfp Tripart
D124 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP132 Weak promoter - ECFP
D104 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP133 Weak promoter - gfp
D104 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon reporter.png
MP134 pLas - ECFP
D105 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP135 pLas - gfp Tripart
D105 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP136 yfp - Double Terminator
D125 (FV) - D117 (FI) 		Part icon reporter.pngPart icon terminator.pngPart icon terminator.png
MP137 rfp - Double Terminator
D125 (FV) - D121 (FI) 		Part icon reporter.pngPart icon terminator.pngPart icon terminator.png
MP138 lasR activator + LVA - Double Terminator
D125 (FV) - D113 (FI) 		Icon coding.pngPart icon terminator.pngPart icon terminator.png
MP139 Strong promoter - gfp tripart
D106 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP140 Strongest RBS - rfp
D102 (BV) - D122 (BI) 		Part icon rbs.pngPart icon reporter.png
MP141 Strongest RBS - yfp
D102 (BV) - D118 (BI) 		Part icon rbs.pngPart icon reporter.png

Glycerol Stock

Protocol

  • 1mL glycerol
  • 1mL culture in LB


List of Strain stocked

Strain Ligation Biobricks Description
S123 L102 Strong rbs - YFP
D129 (BV) - D118 (BI) 		Part icon rbs.pngPart icon reporter.png
S124 L103 Strong rbs - mRFP
D129 (BV) - D122 (BI) 		Part icon rbs.pngPart icon reporter.png
S125 L104 Strong rbs - lasR activator
D129 (BV) - D114 (BI) 		Part icon rbs.pngIcon coding.png
S126 L105 Strong promoter - ECFP
D123 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
S127 L106 Strong promoter - gfp Tripart
D123 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
S128 L107 Strongest promoter - gfp Tripart
D103 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
S129 L108 Weak promoter - ECFP
D103 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
S130 L110 Medium promoter - gfp
D124 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon reporter.png
S131 L111 Weak promoter - ECFP
D104 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
S132 L112 Weak promoter - gfp Tripart
D104 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
S133 L115 pLas - ECFP
D105 (BV) - D130 (BI) 		Part icon reporter.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
S134 L116 pLas - gfp Tripart
D105 (BV) - D131 (BI) 		Part icon reporter.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
S135 L117 yfp - Double Terminator
D125 (FV) - D117 (FI) 		Part icon reporter.pngPart icon terminator.pngPart icon terminator.png
S136 L118 rfp - Double Terminator
D125 (FV) - D117 (FI) 		Part icon reporter.pngPart icon terminator.pngPart icon terminator.png
S137 L119 LasR activator + LVA - Double terminator
D125 (FV) - D113 (FI) 		Icon coding.pngPart icon terminator.pngPart icon terminator.png
S138 L121 Strong promoter - gfp Tripart
D106 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
S139 L124 Strongest rbs - mRFP
D102 (BV) - D122 (BI) 		Part icon rbs.pngPart icon reporter.png
S140 L125 Strongest rbs - yfp
D102 (BV) - D118 (BI) 		Part icon rbs.pngPart icon reporter.png

Transformations

Protocol

use of TOP10 Chemically competent cells

  • Defroze competent cells on ice during 5'
  • Add 5µl of DNA Ligation in 50µL of competent bacterias (or 1µL for the positive control puc19)
  • Incubate 30' on ice
  • Heat-shock the cells during 30" at 42°C without shaking
  • Put 2' on ice
  • Add 250µL of pre-warmed SOC medium (4°C)
  • Incubate 1h at 37°C under shaking (225rpm)
  • Spin at 5.000rpm during 30"
  • Remove 150µL of supernatant
  • Resuspent the pellet in the 150µL left
  • Spread on adequated plates
  • Incubate O/N at 37°C


List of transformation

Name Description Biobrick
L100 RBS-TetR-ECFP Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
L101 RBS-tetR-GFP tripart Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
L113 pBAD-ECFP Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
L114 pBAD-GFP tripart Part icon regulatory.pngPart icon reporter.png
L120 ptet-ECFP Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
L122 RBS-lasI-ECFP Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
L123 RBS-lasI-GFP-ter Part icon rbs.pngIcon coding.pngPart icon reporter.pngPart icon terminator.png
L126 RBS-lasR-LVA Part icon rbs.pngIcon coding.png
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