Team:Paris/July 30

From 2008.igem.org

(Difference between revisions)
(→Results of transformations)
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D110
D110
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D129
D129
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D123
D123
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D103
D103
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D124
D124
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D104
D104
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D126
D126
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D105
D105
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D125
D125
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D106
D106
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D107
D107
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D102
D102
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puc19
puc19
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|align="center"|155 (transformation efficiency:1.5*10^7/ug)
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==Analysis of yesterday DNA digestion==
==Analysis of yesterday DNA digestion==

Revision as of 09:32, 5 August 2008

← Yesterday

↓ Calendar ↑

Tomorrow →


Results of transformations

Ligation name Description Antibio Nb colonies observed Comments
L100

rbs TetR - ECFP
D110 (BV) - D130 (BI)

Amp 0 After another night nothing was osberved -> To do again
L101

rbs TetR - GFP tripart
D110 (BV) - D131 (BI)

Amp 1 After another night nothing was observed -> To do again
L102

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

Amp +/- 100
L103

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

Amp a lot
L104

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

Amp a lot
L105

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

Amp a lot
L106

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

Amp a lot
L107

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

Amp a lot
L108 n°2 (the right one)

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

Amp +/- 100
L109 n°1

Strong promoter - ecfp
D124 (BV) - D130 (BI)

Amp +/- 30 To do again
L109 n°2

Strong promoter - ecfp
D124 (BV) - D130 (BI)

Amp a lot To do again
L110

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

Amp a lot
L111

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

Amp a lot
L112

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

Amp a lot
L113

AracpBAD - ecfp
D126 (BV) - D130 (BI)

Kan 1 After another night nothing was observed -> To do again
L114

AracpBAD - gfp tripart
D126 (BV) - D131 (BI)

Kan 2 After another night nothing was observed -> To do again
L115

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

Amp a lot
L116

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

Amp a lot
L117

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

Amp +/- 30
L118

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

Amp +/- 30
L119

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

Amp +/- 30
L120

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

Amp a lot
L121

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

Amp a lot
L122

RBS-lasI - ecfp
D107 (BV) - D130 (BI)

Amp 0 After another night nothing was observed -> To do again
L123

RBS lasI - ecfp
D107 (BV) - D131 (BI)

Amp 1 After another night nothing was observed -> To do again
L124

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

Amp a lot
L125

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

Amp a lot
L126

Strongest RBS (1)- LacR activator (+LVA)
D102 (BV) - D118 (BI)

Amp no dish found!!! To do again
L127

gfp (1)- Double terminator
D119 (FI) - D125 (FV)

Amp no dish found!!! To do again
Controls
C1

D110

Amp 0 -
C2

D129

Amp 5 -
C3

D123

Amp +/- 100 -
C4

D103

Amp +/- 20 -
C5

D124

Amp +/- 100 -
C6

D104

Amp +/- 100 -
C7

D126

Kana 0 -
C8

D105

Amp +/- 20 -
C9

D125

Amp 0 -
C10

D106

Amp 0 -
C11

D107

Amp 0 -
C12

D102

Amp +/- 10 -
Positive control

puc19

Amp 155 (transformation efficiency:1.5*10^7/ug) -

Analysis of yesterday DNA digestion

The digested DNA of yesterday was analysed one more time by electrophoresis on a 0.8% agarose gel (about 30 minutes at 100 W). The ladder used was the 1 kb DNA ladder (New England Biolabs). 5 µL of each sample with 1 µL of loading dye were loaded.

KR000082.jpg


Name BioBrick Tube N° Enz 1 Enz 2 Obs Exp Size Mea Size Conc (ng/µl) Band
D101 B0034 1 EcoRI XbaI FV 2076 bp 2000 bp 6 10
D102 B0034 1 SpeI PstI BV 2077 bp 2000 bp 8 11
D105 R0079 1 SpeI PstI BV 2222 bp 2000 bp 6 13
D106 R0040 1 SpeI PstI BV 2119 bp 2000 bp 6 12
D108 S03154 1 XbaI PstI BI 707 bp 0 (failed) 14
D111 S03879 1 XbaI PstI BI 725 bp 600 bp 2 15
D115 C0179 2 EcoRI SpeI FI 746 bp 0 (failed) 4 & 5
D116 C0179 2 XbaI PstI BI 745 bp 700 bp 2 2 & 3
D117 E0030 1 EcoRI SpeI FI 746 bp 600 bp 10 16
D128 B0030 2 EcoRI XbaI FV 2079 bp 2000 bp 8 6 & 7
D129 B0030 2 SpeI PstI BV 2080 bp 2000 bp 8 8 & 9

Results : Each of the samples was succesfully digested and purified except for the sample D108. It seems that the QIAprep columms (from the QIAGEN Minipreps kit) can be used instead of the QIAquick columms (for DNA Gel Extraction).

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