Team:Paris/August 18

From 2008.igem.org

(Difference between revisions)
(→Culture of stable strains with biobrocks 2008)
(→Culture of stable strains with biobrocks 2008)
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|'''Description'''
|'''Description'''
|-
|-
-
|[http://partsregistry.org/Part:BBa_B0032 B0032] in [http://partsregistry.org/Part:BBa_pSB1A2 pSB1A2]
+
|[http://partsregistry.org/Part:BBa_B0032 B0032] in [http://partsregistry.org/Part:pSB1A2]
|rbs (efficiency=0.3)
|rbs (efficiency=0.3)
|[[Image:Icon_rbs.png]]
|[[Image:Icon_rbs.png]]
|-
|-
-
|[http://partsregistry.org/Part:BBa_E0422 E0422]
+
|[http://partsregistry.org/Part:BBa_E0422 E0422] in [http://partsregistry.org/Part:pSB1A2]
|ECFP (LVA+)
|ECFP (LVA+)
|[[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
|[[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
|-
|-
-
|[http://partsregistry.org/Part:BBa_E0420 E0420]
+
|[http://partsregistry.org/Part:BBa_E0420 E0420] in [http://partsregistry.org/Part:pSB1A2]
|ECFP (LVA-)
|ECFP (LVA-)
|[[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
|[[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
|-
|-
-
|[http://partsregistry.org/Part:BBa_E0430 E0430]
+
|[http://partsregistry.org/Part:BBa_E0430 E0430] in [http://partsregistry.org/Part:pSB1A2]
|EYFP (LVA-)
|EYFP (LVA-)
|[[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
|[[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
|-
|-
-
|[http://partsregistry.org/Part:BBa_E0432 E0432]
+
|[http://partsregistry.org/Part:BBa_E0432 E0432] in [http://partsregistry.org/Part:pSB1A2]
|EYFP (LVA+)
|EYFP (LVA+)
|[[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
|[[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
|-
|-
-
|[http://partsregistry.org/Part:BBa_I732078 I732078]
+
|[http://partsregistry.org/Part:BBa_I732078 I732078] in [http://partsregistry.org/Part:pSB1A3]
|mRFP1 (LVA+)
|mRFP1 (LVA+)
|[[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
|[[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
|-
|-
-
|[http://partsregistry.org/Part:BBa_I13507 I13507]
+
|[http://partsregistry.org/Part:BBa_I13507 I13507] in [http://partsregistry.org/Part:pSB1A2]
|mRFP1 (LVA-)
|mRFP1 (LVA-)
|[[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
|[[Image:Icon_rbs.png]][[Image:Icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
|}
|}

Revision as of 16:07, 19 August 2008

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Contents

Analysis of the cloning of OmpR and EnvZ

PCR screening

  • positive control: S158 (pSB3K3)
  • negative control: no template
  • screening PCR programm
  • elongation time: 1 min 30
  • primers: O18 & O19

Reaction mixture (25 µL total volume)

  • 12,5 µL Quick load PCR Mix
  • 0,5 µL O18
  • 0,5 µL O19
  • 12 µL water

We wanted to analyse the transformants of the ligations L133 (OmpR in pSB1A2) and L134 (EnvZ in pSB1A2) from the experience of august 9th/11th.
3 clones of L133 and 3 clones of L134 were analysed.
Each clone tested was then cultured in 7,5 mL LB + ampicilline.


Miniprep and stock glycerol of L151

Stock number Ligation name Biobricks Description
S162.1 L151.1 Part icon rbs.pngIcon coding.pngPart icon terminator.pngPart icon terminator.png rbs-LasR - Double terminator
S162.2 L151.2
S162.3 L151.3


Miniprep Name Ligation name Antibio Biobricks Description
MP162.1 L151.1 Kan Part icon rbs.pngIcon coding.pngPart icon terminator.pngPart icon terminator.png rbs-LasR - Double terminator
MP162.2 L151.2
MP162.3 L151.3

Digestion of MP162

Measurement of the concentration of MPs 162

Miniprep Name Concentration (µg/mL) Ratio 260/280
MP162.1 151 1.71
MP162.2 172 1.73
MP162.3 149 1.73

Digestion

Protocol

Digestion name Template DNA Enzymes Volume of DNA
D165 MP162.2 XbaI - PstI 5.81 µL

Ligation

Protocol

Ligation Name Vector Name Volume Vector (µL) Insert Volume Insert (µL)
L153 D123.1 1.54 D165 1.69
L154 D103.2 1.14 D165 1.68

Culture of stable strains with biobrocks 2008

Protocol

Biobrick Name Part Description
B0032 in [1] rbs (efficiency=0.3) Icon rbs.png
E0422 in [2] ECFP (LVA+) Icon rbs.pngIcon reporter.pngPart icon terminator.pngPart icon terminator.png
E0420 in [3] ECFP (LVA-) Icon rbs.pngIcon reporter.pngPart icon terminator.pngPart icon terminator.png
E0430 in [4] EYFP (LVA-) Icon rbs.pngIcon reporter.pngPart icon terminator.pngPart icon terminator.png
E0432 in [5] EYFP (LVA+) Icon rbs.pngIcon reporter.pngPart icon terminator.pngPart icon terminator.png
I732078 in [6] mRFP1 (LVA+) Icon rbs.pngIcon reporter.pngPart icon terminator.pngPart icon terminator.png
I13507 in [7] mRFP1 (LVA-) Icon rbs.pngIcon reporter.pngPart icon terminator.pngPart icon terminator.png
Retrieved from "http://2008.igem.org/Team:Paris/August_18"