Paris/August 22

From 2007.igem.org

(Difference between revisions)
(→Growth kinetic on w121 strains)
(→Minipreps of Ligation transformation strains from yesterday)
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== Minipreps of Ligation transformation strains from yesterday ==
== Minipreps of Ligation transformation strains from yesterday ==
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== Plasmids: MiniPrep Products ==
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{|
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|- style="background: #ccccff;"
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! colspan="5" style="background: #ccccff;" | Plasmids: MiniPrep Products
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|- style="background: #ccccff; text-align: center;"
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|width=5%| Number
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|width=25%| Name
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|width=50%| Description
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|width=15%| Products in use
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|width=5%| Date
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|- style="background: #cccccc;" 
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| style="background: #ccffcc;" |L31
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|<bbpart>BBa_I0500</bbpart> in <bbpart>J61002</bbpart>
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|construct with pBad promotor regulating expression of mRFP <partinfo>I0500 SpecifiedComponents</partinfo>
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| L31.1, L31.3, L31.4
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| [[Paris/August 20| August 20]]
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|- style="background: #cccccc;" 
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| style="background: #ccffcc;" |L32
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|Lox66DapAE.coli in <bbpart>pSB1A2</bbpart>
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|Insertion of lox66DapAE.coli construct as a biobrick in pSB1A2 plasmid
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http://partsregistry.org/wiki/images/9/9f/Part_icon_dna.png
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http://partsregistry.org/wiki/images/7/7b/Part_icon_cds.png
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|L32.1, L32.2, L32.3
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| [[Paris/August 20| August 20]]
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|- style="background: #cccccc;" 
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| style="background: #ccffcc;" |L34
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|<bbpart>BBa_B0030</bbpart>+<bbpart>BBa_J61047</bbpart>
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|strong RBS(B0030)-Cre ORF
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http://partsregistry.org/images/partbypart/icon_rbs.png
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http://partsregistry.org/wiki/images/7/7b/Part_icon_cds.png
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|L34.1, L34.2, L34.3
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| [[Paris/August 20| August 20]]
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|- style="background: #cccccc;" 
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| style="background: #ccffcc;" |L35
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|lox66 in <bbpart>pSB1A2</bbpart>
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|
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|L35.1, L35.2, L35.3
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| [[Paris/August 20| August 20]]
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|- style="background: #cccccc;" 
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| style="background: #ccffcc;" |L36
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|lox71 in <bbpart>pSB1A2</bbpart>
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|
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|L36.1, L36.2, L36.3
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| [[Paris/August 20| August 20]]
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|- style="background: #cccccc;" 
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| style="background: #ccffcc;" |L37
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|lox71-gfp in pSB1A2
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|construction of the biobrick lox71-gfp
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http://partsregistry.org/wiki/images/9/9f/Part_icon_dna.png
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http://partsregistry.org/images/partbypart/icon_reporter.png
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|L37.1, L37.2, L37.3
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| [[Paris/August 20| August 20]]
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|- style="background: #cccccc;" 
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| style="background: #ccffcc;" |L39
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|araC/pbad>>RBS-Cre in <bbpart>J61002</bbpart>
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|ligation ([[Paris/August 13| Aug.13]]) of RBS-Cre as a CDS behind pbad in <bbpart>J61002</bbpart>
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|L39.1, L39.2, L39.3
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| [[Paris/August 20| August 20]]
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|- style="background: #cccccc;" 
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| style="background: #ccffcc;" |L40
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|araC/pbad>>ftsZ <bbpart>J61002</bbpart>
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|ligation ([[Paris/August 13| Aug.13]]) of ftsZ as a CDS behind pbad in <bbpart>J61002</bbpart>
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|L40.1, L40.2
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| [[Paris/August 20| August 20]]
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|- style="background: #cccccc;" 
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| style="background: #ccffcc;" |L41
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|araC/pbad>>ftsA-ftsZ <bbpart>J61002</bbpart>
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|ligation ([[Paris/August 13| Aug.13]]) of ftsA-Z as a CDS behind pbad in <bbpart>J61002</bbpart>
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|L41.1
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| [[Paris/August 20| August 20]]
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|- style="background: #cccccc;"
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| style="background: #ccffcc;" |L42
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|lox66-gfptripart <bbpart>pSB1A2</bbpart>
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|ligation ([[Paris/August 13| Aug.13]]) of gfp-tripart behind lox66
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|L42.1, L42.2, L42.3
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| [[Paris/August 20| August 20]]
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|- style="background: #cccccc;"
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| style="background: #ccffcc;" |L43
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|dapAp>>mRFP in <bbpart>J61002</bbpart>
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|dapAp promotor placed([[Paris/August 13| Aug.13]]) upstream of mRFP <bbpart>J61002</bbpart>
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|L43.1, L43.2, L43.3
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| [[Paris/August 20| August 20]]
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|}
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*L31.1 - .3 - .4
*L31.1 - .3 - .4

Revision as of 09:12, 22 August 2007

yesterday -- tomorrow

Contents

Minipreps of Ligation transformation strains from yesterday

Plasmids: MiniPrep Products

Plasmids: MiniPrep Products
Number Name Description Products in use Date
L31 BBa_I0500 in J61002 construct with pBad promotor regulating expression of mRFP L31.1, L31.3, L31.4 August 20
L32 Lox66DapAE.coli in pSB1A2 Insertion of lox66DapAE.coli construct as a biobrick in pSB1A2 plasmid

http://partsregistry.org/wiki/images/9/9f/Part_icon_dna.png http://partsregistry.org/wiki/images/7/7b/Part_icon_cds.png

L32.1, L32.2, L32.3 August 20
L34 BBa_B0030+BBa_J61047 strong RBS(B0030)-Cre ORF

http://partsregistry.org/images/partbypart/icon_rbs.png http://partsregistry.org/wiki/images/7/7b/Part_icon_cds.png

L34.1, L34.2, L34.3 August 20
L35 lox66 in pSB1A2 L35.1, L35.2, L35.3 August 20
L36 lox71 in pSB1A2 L36.1, L36.2, L36.3 August 20
L37 lox71-gfp in pSB1A2 construction of the biobrick lox71-gfp

http://partsregistry.org/wiki/images/9/9f/Part_icon_dna.png http://partsregistry.org/images/partbypart/icon_reporter.png

L37.1, L37.2, L37.3 August 20
L39 araC/pbad>>RBS-Cre in J61002 ligation ( Aug.13) of RBS-Cre as a CDS behind pbad in J61002 L39.1, L39.2, L39.3 August 20
L40 araC/pbad>>ftsZ J61002 ligation ( Aug.13) of ftsZ as a CDS behind pbad in J61002 L40.1, L40.2 August 20
L41 araC/pbad>>ftsA-ftsZ J61002 ligation ( Aug.13) of ftsA-Z as a CDS behind pbad in J61002 L41.1 August 20
L42 lox66-gfptripart pSB1A2 ligation ( Aug.13) of gfp-tripart behind lox66 L42.1, L42.2, L42.3 August 20
L43 dapAp>>mRFP in J61002 dapAp promotor placed( Aug.13) upstream of mRFP J61002 L43.1, L43.2, L43.3 August 20


  • L31.1 - .3 - .4
  • L32.1 - .2 - .3
  • L34.1 - .2 - .3
  • L35.1 - .2 - .3
  • L36.1 - .2 - .3
  • L37.1 - .2 - .3
  • L39.1 - .2 - .3
  • L40.1 - .2
  • L41.1
  • L42.1 - .2 - .3
  • L43.1 - .2 - .3

Growth kinetic on w121 strains

In order to precise the growth behavior/kinetics of our strain W121 in smaller DAP concentration, we fit those last ones in function of our last kinetics results (06/08/07)

Kinetic Array :w121 kinetic as a function of DAP and supplemented medium (S0.x)
1 2 3 4 5 6 7 8 9 10 11 12
A H20 H20 H20 H20 H20 H20 H20 H20 H20 H20 H20 H20
B H20 LB+0µM DAP LB+5µM DAP LB+8µM DAP LB+10µM DAP LB+12µM DAP LB+15µM DAP LB+17µM DAP LB+20µM DAP LB+30µM DAP LB+40µM DAP H20
C H20 S0.2+0µM DAP S0.2+5µM DAP S0.2+6µM DAP S0.2+7µM DAP S0.2+8µM DAP S0.2+9µM DAP S0.2+10µM DAP S0.2+15µM DAP S0.2+17µM DAP S0.2+20µM DAP H20
D H20 S0.4+0µM DAP S0.4+2µM DAP S0.4+3µM DAP S0.4+4µM DAP S0.4+5µM DAP S0.4+8µM DAP S0.4+10µM DAP S0.4+12µM DAP S0.4+15µM DAP S0.4+20µM DAP H20
E H20 S0.6+0µM DAP S0.6+1µM DAP S0.6+2µM DAP S0.6+3µM DAP S0.6+4µM DAP S0.6+5µM DAP S0.6+6µM DAP S0.6+7µM DAP S0.6+8µM DAP S0.6+10µM DAP H20
F H20 S0.8+0µM DAP S0.8+0,5µM DAP S0.8+1µM DAP S0.8+2µM DAP S0.8+3µM DAP S0.8+4µM DAP S0.8+5µM DAP S0.8+6µM DAP S0.8+7µM DAP S0.8+10µM DAP H20
G H20 H20 H20 H20 H20 H20 H20 H20 H20 H20 H20 H20
H H20 H20 H20 H20 H20 H20 H20 H20 H20 H20 H20 H20

The 96 well plate is surrounded by H2O in order to maintain humidity during the assay. In each slot, 200µL of the growth medium (LB or S0.x) is mixed with 2µL of w121 culture grown ON, and with different amount of DAP (see table). The growth profile is measured over a period of 20H, a DO measurement is acquired each 4min10s.

Preparing chemically competent cells

  • w121
  • FX85

Transformation of W121 and FX85 strains

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