Team:Paris/July 31

From 2008.igem.org

(Difference between revisions)
Line 163: Line 163:
=== Protocol ===
=== Protocol ===
 +
 +
For each samples,
 +
 +
* 1 µl Ligase
 +
* X µl Vector
 +
* Y µl Insert (3x more than vector)
 +
* 2 µl Ligase Buffer 10x
 +
* 20 µl qsp H2O
 +
 +
 +
* Incubates O/N at 4°C (in the fridge)
 +
 +
 +
 +
{| border="1"
 +
|align=center|'''Name'''
 +
|align=center|'''Vector'''
 +
|align=center|'''Insert'''
 +
|align=center|'''Antibio'''
 +
|align=center|'''Vol. vector (µl)'''
 +
|align=center|'''Vol. insert (µl)'''
 +
|align=center|'''Vol. H20 (µl)'''
 +
|-
 +
|style="background: #ccccff;"| L100
 +
|align=center| D110
 +
|align=center| D130
 +
|align=center| A
 +
|align=center|3
 +
|align=center|6
 +
|align=center|8
 +
|-
 +
|style="background: #ccccff;"| L101
 +
|align=center| D110
 +
|align=center| D131
 +
|align=center| A
 +
|align=center|3
 +
|align=center|6
 +
|align=center|8
 +
|-
 +
|style="background: #ccccff;"|  L109
 +
|align=center| D124
 +
|align=center| D130
 +
|align=center| A
 +
|align=center|4
 +
|align=center|7
 +
|align=center|6
 +
|-
 +
|style="background: #ccccff;"|  L113
 +
|align=center| D126
 +
|align=center| D130
 +
|align=center| K
 +
|align=center|5
 +
|align=center|2.5
 +
|align=center|9.5
 +
|-
 +
|style="background: #ccccff;"|  L114
 +
|align=center| D126
 +
|align=center| D131
 +
|align=center| K
 +
|align=center|5
 +
|align=center|2.5
 +
|align=center|9.5
 +
|-
 +
|style="background: #ccccff;"|  L120
 +
|align=center| D106
 +
|align=center| D130
 +
|align=center| A
 +
|align=center|6
 +
|align=center|7.5
 +
|align=center|3.5
 +
|-
 +
|style="background: #ccccff;"|  L122
 +
|align=center| D107
 +
|align=center| D130
 +
|align=center| A
 +
|align=center|7
 +
|align=center|3.5
 +
|align=center|6.5
 +
|-
 +
|style="background: #ccccff;"| L123
 +
|align=center| D107
 +
|align=center| D131
 +
|align=center| A
 +
|align=center|7
 +
|align=center|3.5
 +
|align=center|6.5
 +
|-
 +
|style="background: #ccccff;"|  L126
 +
|align=center| D102
 +
|align=center| D114
 +
|align=center| A
 +
|align=center|6
 +
|align=center|4
 +
|align=center|7
 +
|-
 +
|style="background: #ccccff;"|  L127
 +
|align=center| D125
 +
|align=center| D119
 +
|align=center| A
 +
|align=center|6
 +
|align=center|3
 +
|align=center|8
 +
|-
 +
|}

Revision as of 10:06, 5 August 2008

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Results of the transformation : Fluorescence

Ligation name Description Fluorescence Comments
L102

Strong rbs - YFP
D129 (BV) - D118 (BI)

No OK
L103

Strong rbs - GFP
D129 (BV) - D122 (BI)

No OK
L104

Strong rbs - lasR activator
D129 (BV) - D114 (BI)

No OK
L105

Strong promoter - ECFP
D123 (BV) - D130 (BI)

Yes (some others red because of the promoter) OK
L106

Strong promoter - gfp Tripart
D123 (BV) - D131 (BI)

Yes (some others red because of the promoter) OK
L107

Strongest promoter - ECFP
D103 (BV) - D131 (BI)

Yes (some others red because of the promoter) OK
L108 n°2 (the right one)

Strong promoter - gfp Tripart
D103 (BV) - D130 (BI)

Yes (some others red because of the promoter) OK
L110

Medium promoter - gfp Tripart
D124 (BV) - D131 (BI)

Yes (some others red because of the promoter) OK
L111

Weak promoter - ECFP
D104 (BV) - D130 (BI)

No To do again
L112

Weak promoter - gfp
D104 (BV) - D131 (BI)

No To do again
L115

pLas - ECFP
D105 (BV)- D130 (BI)

No OK
L116

pLas - gfp Tripart
D105 (BV) - D131 (BI)

1 green To do again
L117

yfp - Double Terminator
D117 (FI) - D125 (FV)

No OK
L118

rfp - Double Terminator
D121 (FI) - D125 (FV)

No OK
L119

lasR activator + LVA - Double Terminator
D113 (FI) - D125 (FV)

No OK
L120

tetR repressible - ECFP
D106 (BV) - D130 (BI)

No To do again
L121

Strong promoter - gfp tripart
D106 (BV) - D130 (BI)

Yes (some others red because of the promoter) OK
L124

Strongest RBS - rfp
D102 (BV) - D122 (BI)

No OK
L125

Strongest RBS - yfp
D102 (BV) - D118 (BI)

No OK


Ligations

We tried again the following ligations : L100 L101 L109 L113 L114 L120 L122 L123 L126 L127

Protocol

For each samples,

  • 1 µl Ligase
  • X µl Vector
  • Y µl Insert (3x more than vector)
  • 2 µl Ligase Buffer 10x
  • 20 µl qsp H2O


  • Incubates O/N at 4°C (in the fridge)


Name Vector Insert Antibio Vol. vector (µl) Vol. insert (µl) Vol. H20 (µl)
L100 D110 D130 A 3 6 8
L101 D110 D131 A 3 6 8
L109 D124 D130 A 4 7 6
L113 D126 D130 K 5 2.5 9.5
L114 D126 D131 K 5 2.5 9.5
L120 D106 D130 A 6 7.5 3.5
L122 D107 D130 A 7 3.5 6.5
L123 D107 D131 A 7 3.5 6.5
L126 D102 D114 A 6 4 7
L127 D125 D119 A 6 3 8
Retrieved from "http://2008.igem.org/Team:Paris/July_31"