Team:Paris/August 13

From 2008.igem.org

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| rowspan="3"| Strongest RBS (1)- LasR activator (+LVA)<br>D102 (BV) - D114 (BI)
| rowspan="3"| Strongest RBS (1)- LasR activator (+LVA)<br>D102 (BV) - D114 (BI)
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Revision as of 08:56, 14 August 2008

← Yesterday

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Contents

Sequencing Minipreps we did yesterday

  • O18 has been diluted to have a concentration of 5µM
  • In each tube : 1µL of O18 diluted
  • Pure water qsp 15µL


Name Ligation Concentration MP (µg/mL) Vol MP (µL) Vol H2O (µL) n° tube
MP144.3 L128.3 163 3.68 10.32 AD1
MP144.4 L128.4 154 3.90 10.10 AD2
MP145.6 L129.6 163 3.68 10.32 AD3
MP145.7 L129.7 188 3.19 10.81 AD4
MP146.1 L130.1 298 2.01 11.99 AD5
MP146.2 L130.2 142 4.23 9.77 AD6

Minipreps: Plasmid extraction

  • Protocol (see # 3) Experiments done by QIAcube
  • List of Minipreps
Name Ligation Biobricks Description
MP147.1 L100.1 rbs TetR - ECFP
D110 (BV) - D130 (BI) 		Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP147.2 L100.2
MP147.3 L100.3
MP148.1 L101.1 rbs TetR - GFP tripart
D110 (BV) - D131 (BI) 		Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP148.2 L101.2
MP148.3 L101.3
MP149.1 L114.1 AracpBAD - gfp tripart
D126 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP149.2 L114.2
MP149.3 L114.3
MP150.1 L120.1 tetR repressible promoter - ECFP
D106 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP150.2 L120.2
MP150.3 L120.3
MP151.1 L122.1 RBS-lasI - ECFP
D107 (BV) - D130 (BI) 		Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP152.1 L123.1 RBS lasI - gfp tripart
D107 (BV) - D131 (BI) 		Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
MP151.2 L123.2
MP152.3 L123.3
MP153.1 L126.1 Strongest RBS (1)- LasR activator (+LVA)
D102 (BV) - D114 (BI) 		Part icon rbs.pngIcon coding.png
MP153.2 L126.2
MP153.3 L126.3
MP154.1 L132.1 pSB1A2 - flhDC
D142(FV) - D139(FI) 		Icon plasmid.pngIcon coding.png
MP154.2 L132.2
MP154.3 L132.3
MP155.1 L133.1 p1SB1A2 - OmpR*
D142(FV) - D140(FI) 		Icon plasmid.pngIcon coding.png
MP156.1 L134.1 p1SB1A2 - EnvZ*
D142(FV) - D141(FI) 		Icon plasmid.pngIcon coding.png
MP156.2 L134.2
MP156.3 L134.3
MP157.1 L138.1 pSB3K3 - gfp generator (E0240)
D137(FV) - D138(FI) 		Icon plasmid.pngIcon rbs.pngIcon reporter.pngPart icon terminator.png

Glycerol Stocks

  • List of Stocks
Strain Ligation Biobricks Description
S146.1 L100.1 rbs TetR - ECFP
D110 (BV) - D130 (BI) 		Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
S146.2 L100.2
S146.3 L100.3
S147.1 L101.1 rbs TetR - GFP tripart
D110 (BV) - D131 (BI) 		Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
S147.2 L101.2
S147.3 L101.3
S148.1 L114.1 AracpBAD - gfp tripart
D126 (BV) - D131 (BI) 		Part icon regulatory.pngPart icon reporter.png
S148.2 L114.2
S148.3 L114.3
S149.1 L120.1 tetR repressible promoter - ECFP
D106 (BV) - D130 (BI) 		Part icon regulatory.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
S149.2 L120.2
S149.3 L120.3
S150.1 L122.1 RBS-lasI - ECFP
D107 (BV) - D130 (BI) 		Part icon rbs.pngIcon coding.pngPart icon rbs.pngPart icon reporter.pngPart icon terminator.png
S151.1 L123.1 RBS lasI - ECFP
D107 (BV) - D131 (BI) 		Part icon rbs.pngIcon coding.pngPart icon reporter.pngPart icon terminator.png
S151.2 L123.2
S151.3 L123.3
S152.1 L126.1 Strongest RBS (1)- LasR activator (+LVA)
D102 (BV) - D114 (BI) 		Part icon rbs.pngIcon coding.png
S152.2 L126.2
S152.3 L126.3

PCR screening

4 more transformants of L130 (pFlhB into J61002) are screened by PCR.

  • PCR screening programm
  • elogation time : 1 min 30
  • template: colonies from the transformation plate
  • positive control: S142 (J61002)
  • negative control: no template
  • primers: O18 and O19



Promoter Characterization: work plan
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