Team:Paris/July 25

From 2008.igem.org

(Difference between revisions)
(→Extraction of DNA from gel)
(→MiniPreps / Digestion)
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==> we decide to repeat all the samples from the beginning, so we culture O/N at 37°c all the strains usefull for the experiments
==> we decide to repeat all the samples from the beginning, so we culture O/N at 37°c all the strains usefull for the experiments
-
== MiniPreps / Digestion ==
+
== MiniPreps ==
-
* Minipreps : Protocol of Promega has been done on all the clones cultivated on the 24th.
 
 +
* Use of Promega's protocol on all the clones cultivated on the 24th.
 +
* Preparation of 50µl of minipreps in simple.
-
===Digestion Mix===
 
-
10µl of Miniprep <br>
 
-
12.5µl of water<br>
 
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2.5µl of Buffer N°2<br>
 
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0.25µl of BSA 100x<br>
 
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1µl of enzyme 1<br>
 
-
1µl of enzyme 2<br>
 
-
 
+
===list of Minipreps===
-
===list of digestion===
+
{| border="1"  
{| border="1"  
|align="center"|'''Name'''
|align="center"|'''Name'''
-
|align="center"|'''Miniprep'''
 
|align="center"|'''Biobrick'''
|align="center"|'''Biobrick'''
-
|align="center"|'''Enzyme 1'''
+
|align="center"|'''Description'''
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|align="center"|'''Enzyme 2'''
+
-
|align="center"|'''Nber tube'''
+
|-
|-
-
|align="center"|D100
 
|align="center"|MP100
|align="center"|MP100
-
|align="center"|B0034
+
|align="center"|[http://partsregistry.org/Part:BBa_B0034 B0034]
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|align="center"|XbaI
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|align="center"|Strongest RBS (Efficiency = 1)
-
|align="center"|PstI
+
-
|align="center"|2
+
|-
|-
-
|align="center"|
 
-
|align="center"|
 
-
|align="center"|
 
-
|align="center"|EcoRI
 
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|align="center"|XbaI
 
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|align="center"|1
 
-
|-
 
-
|align="center"|
 
-
|align="center"|
 
-
|align="center"|
 
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|align="center"|SpeI
 
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|align="center"|PstI
 
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|align="center"|2
 
-
|-
 
-
|align="center"|D116
 
|align="center"|MP116
|align="center"|MP116
-
|align="center"|J23100
+
|align="center"|[http://partsregistry.org/Part:BBa_J23100 J23100]
-
|align="center"|SpeI
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|align="center"|Strong constitutive promoter in J61002
-
|align="center"|PstI
+
-
|align="center"|2
+
|-
|-
-
|align="center"|D120
 
|align="center"|MP120
|align="center"|MP120
-
|align="center"|B0030
+
|align="center"|[http://partsregistry.org/Part:BBa_B0030 B0030]
-
|align="center"|SpeI
+
|align="center"|Strong RBS (Efficiency = 0,6)
-
|align="center"|PstI
+
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|align="center"|2
+
|-
|-
-
|align="center"|
 
-
|align="center"|
 
-
|align="center"|
 
-
|align="center"|EcoRI
 
-
|align="center"|XbaI
 
-
|align="center"|1
 
-
|-
 
-
|align="center"|
 
-
|align="center"|
 
-
|align="center"|
 
-
|align="center"|XbaI
 
-
|align="center"|PstI
 
-
|align="center"|2
 
-
|-
 
-
|align="center"|D121
 
|align="center"|MP121
|align="center"|MP121
-
|align="center"|E0422
+
|align="center"|[http://partsregistry.org/Part:BBa_E0422 E0422]
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|align="center"|XbaI
+
|align="center"|ECFP (RBS+LVA+Term)
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|align="center"|PstI
+
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|align="center"|1 !
+
-
|-
+
-
|align="center"|D122
+
-
|align="center"|MP122
+
-
|align="center"|E0840
+
-
|align="center"|XbaI
+
-
|align="center"|PstI
+
-
|align="center"|1 !
+
-
|}
+
-
 
+
-
 
+
-
=== Results of digestions : Electrophoresis ===
+
-
 
+
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{| border="1"
+
-
|align=center|'''Name'''
+
-
|align=center|'''BioBrick'''
+
-
|align=center|'''Tube N°'''
+
-
|align=center|'''Enz 1'''
+
-
|align=center|'''Enz 2'''
+
-
|align=center|'''Obs'''
+
-
|align=center|'''Exp Size Matrix'''
+
-
|align=center|'''Exp Size BB'''
+
-
|align=center|'''Mea Size Matrix'''
+
-
|align=center|'''Mea Size BB'''
+
-
|-
+
-
! rowspan="5" style="background: #ccccff;" | D100
+
-
! rowspan="5" style="background: #ccccff;" | B0034
+
-
|align=center|1
+
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! rowspan="2"| XbaI
+
-
! rowspan="2"| PstI
+
-
! rowspan="2"| BI
+
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! rowspan="2"| 2057 pb
+
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! rowspan="2"| 34 pb
+
-
|align=center|
+
-
|align=center|
+
-
|-
+
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|align=center|2
+
-
|align=center|
+
-
|align=center|
+
-
|-
+
-
|align=center|3
+
-
|align=center| EcoRI
+
-
|align=center| XbaI
+
-
|align=center| FV
+
-
|align=center| 2076 pb
+
-
|align=center| 15 pb
+
-
|align=center|
+
-
|align=center|
+
-
|-
+
-
|align=center|4
+
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! rowspan="2"| SpeI
+
-
! rowspan="2"| PstI
+
-
! rowspan="2"| BV
+
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! rowspan="2"| 2077 pb
+
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! rowspan="2"| 14 pb
+
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|align=center|
+
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|align=center|
+
-
|-
+
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|align=center|5
+
-
|align=center|
+
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|align=center|
+
-
|-
+
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! rowspan="1" style="background: #ccccff;"|D116
+
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! rowspan="1" style="background: #ccccff;"|J23100
+
-
|align=center|1
+
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! rowspan="1"| SpeI
+
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! rowspan="1"| PstI
+
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! rowspan="1"| BV
+
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! rowspan="1"| 2100 pb
+
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! rowspan="1"| 883 pb
+
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|align=center|
+
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|align=center|
+
-
|-
+
-
! rowspan="5" style="background: #ccccff;" | D120
+
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! rowspan="5" style="background: #ccccff;" | B0030
+
-
|align=center|1
+
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! rowspan="2"| XbaI
+
-
! rowspan="2"| PstI
+
-
! rowspan="2"| BI
+
-
! rowspan="2"| 2057 pb
+
-
! rowspan="2"| 37 pb
+
-
|align=center|
+
-
|align=center|
+
-
|-
+
-
|align=center|2
+
-
|align=center|
+
-
|align=center|
+
-
|-
+
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|align=center|3
+
-
|align=center| EcoRI
+
-
|align=center| XbaI
+
-
|align=center| FV
+
-
|align=center| 2079 pb
+
-
|align=center| 15 pb
+
-
|align=center|
+
-
|align=center|
+
-
|-
+
-
|align=center|4
+
-
! rowspan="2"| SpeI
+
-
! rowspan="2"| PstI
+
-
! rowspan="2"| BV
+
-
! rowspan="2"| 2080 pb
+
-
! rowspan="2"| 14 pb
+
-
|align=center|
+
-
|align=center|
+
-
|-
+
-
|align=center|5
+
-
|align=center|
+
-
|align=center|
+
-
|-
+
-
! rowspan="1" style="background: #ccccff;"|D121
+
-
! rowspan="1" style="background: #ccccff;"|E0422
+
-
|1
+
-
|XbaI
+
-
|PstI
+
-
| FV
+
-
| 2057 pb
+
-
| 939 pb
+
-
|
+
-
|
+
|-
|-
-
! rowspan="1" style="background: #ccccff;"|D122
+
|align="center"| MP122
-
! rowspan="1" style="background: #ccccff;"|E0840
+
|align="center"|[http://partsregistry.org/Part:BBa_E0840 E0840]
-
|1
+
|align="center"|gfp tri-part; strong rbs
-
|XbaI
+
-
|PstI
+
-
| FV
+
-
| 2057 pb
+
-
| 900 pb
+
-
|
+
|
|
|}
|}

Revision as of 11:22, 28 July 2008

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Extraction of the DNA

  • Use of Promega's protocol for the extraction of: gel n°1 --> 4 : (D100 --> D110 cl-1 ES))
  • Use of Qiagen's protocol for the extraction of: gel n°5 --> 8 : (D110 cl-2 ES --> D120))
  • Test of the succeed of the extraction by electrophoresis on 2µl of the parts extracted.


==> conclusion : we don't succeed to detect DNA by spectrophometry or electrophoresis for the samples produced by Qiagen's protocol.


==> we decide to repeat all the samples from the beginning, so we culture O/N at 37°c all the strains usefull for the experiments

MiniPreps

  • Use of Promega's protocol on all the clones cultivated on the 24th.
  • Preparation of 50µl of minipreps in simple.


list of Minipreps

Name Biobrick Description
MP100 [http://partsregistry.org/Part:BBa_B0034 B0034] Strongest RBS (Efficiency = 1)
MP116 [http://partsregistry.org/Part:BBa_J23100 J23100] Strong constitutive promoter in J61002
MP120 [http://partsregistry.org/Part:BBa_B0030 B0030] Strong RBS (Efficiency = 0,6)
MP121 [http://partsregistry.org/Part:BBa_E0422 E0422] ECFP (RBS+LVA+Term)
MP122 [http://partsregistry.org/Part:BBa_E0840 E0840] gfp tri-part; strong rbs
Retrieved from "http://2008.igem.org/Team:Paris/July_25"