Team:Paris/September 2

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{{Paris/Calendar_Links|September 1|September 3}}
{{Paris/Calendar_Links|September 1|September 3}}
-
=Ligation L173 (rbs-TetR in mRFP-LVA-tripart-pSB1A3)=
+
=Construction of rbs-TetR-mRFP-LVA-tripart (L173)=
-
 
+
[[Image:Part_icon_rbs.png]][[Image:icon_coding.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
 +
=='''Ligation L173'''==
{|border="1" style="text-align: center"
{|border="1" style="text-align: center"
 +
|'''Digestion n°'''
|'''Name'''
|'''Name'''
-
|'''Digestion number'''
 
|'''Digested by'''
|'''Digested by'''
 +
|'''DNA concentration'''
|-
|-
-
|rbs-TetR
 
|D112
|D112
 +
|rbs-TetR
|EcoRI & SpeI (insert)
|EcoRI & SpeI (insert)
 +
|~20 µg/mL
|-
|-
-
|mRFP-LVA-tripart
 
|D187
|D187
 +
|mRFP-LVA-tripart
|EcoRI & XbaI (vector)
|EcoRI & XbaI (vector)
 +
|16 µg/mL
|-
|-
|}
|}
<br>
<br>
 +
==='''Reaction mixtures'''===
{|border="1" style="text-align: center"
{|border="1" style="text-align: center"
|rowspan="2"|
|rowspan="2"|
 +
|rowspan="2"|'''Control'''
|rowspan="2"|'''Control'''
|colspan="2"|'''Sample'''
|colspan="2"|'''Sample'''
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|'''4:1''' (insert:vector) '''ratio'''
|'''4:1''' (insert:vector) '''ratio'''
|-
|-
-
|D112 (insert)
+
|'''D112''' (insert)
|0 µL
|0 µL
|1,4 µL
|1,4 µL
|1,9 µL
|1,9 µL
|-
|-
-
|D187 (vector)
+
|'''D187''' (vector)
|2,5 µL
|2,5 µL
|2,5 µL
|2,5 µL
|2,5 µL
|2,5 µL
|-
|-
-
|T4 DNA ligase 10X buffer
+
|T4 DNA ligase 10X '''buffer'''
|2 µL
|2 µL
|2 µL
|2 µL
|2 µL
|2 µL
|-
|-
-
|T4 DNA ligase
+
|T4 DNA '''ligase'''
|1 µL
|1 µL
|1 µL
|1 µL
|1 µL
|1 µL
|-
|-
-
|water
+
|'''water'''
|14,5 µL
|14,5 µL
|13,1 µL
|13,1 µL
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|-
|-
|}
|}
 +
* overnight reaction at about 16°C.
 +
<br>
 +
 +
=Construction of rbs-FlhDC-mRFP-LVA-tripart (L172)=
 +
[[Image:Part_icon_rbs.png]][[Image:icon_coding.png]][[Image:Part_icon_reporter.png]][[Image:Part_icon_terminator.png]][[Image:Part_icon_terminator.png]]
 +
 +
=='''Ligation L172'''==
 +
{|border="1" style="text-align: center"
 +
|'''Digestion n°'''
 +
|'''Name'''
 +
|'''Digested by'''
 +
|'''DNA concentration'''
 +
|-
 +
|D186
 +
|rbs-FlhDC
 +
|EcoRI & SpeI (insert)
 +
|~24 µg/mL
 +
|-
 +
|D187
 +
|mRFP-LVA-tripart
 +
|EcoRI & XbaI (vector)
 +
|16 µg/mL
 +
|-
 +
|}
 +
<br>
 +
==='''Reaction mixtures''' ('''3:1''' (insert:vector) '''ratio''')===
 +
{|border="1" style="text-align: center"
 +
|
 +
|'''Control'''
 +
|'''Sample'''
 +
|-
 +
|'''D186''' (insert)
 +
|0 µL
 +
|1 µL
 +
|-
 +
|'''D187''' (vector)
 +
|2,5 µL
 +
|2,5 µL
 +
|-
 +
|T4 DNA ligase 10X '''buffer'''
 +
|2 µL
 +
|2 µL
 +
|-
 +
|T4 DNA '''ligase'''
 +
|1 µL
 +
|1 µL
 +
|-
 +
|'''water'''
 +
|14,5 µL
 +
|13,5 µL
 +
|-
 +
|}
 +
 +
* overnight reaction at about 16°C.

Latest revision as of 19:24, 9 September 2008

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Contents

Construction of rbs-TetR-mRFP-LVA-tripart (L173)

Part icon rbs.pngIcon coding.pngPart icon reporter.pngPart icon terminator.pngPart icon terminator.png

Ligation L173

Digestion n° Name Digested by DNA concentration
D112 rbs-TetR EcoRI & SpeI (insert) ~20 µg/mL
D187 mRFP-LVA-tripart EcoRI & XbaI (vector) 16 µg/mL


Reaction mixtures

Control Sample
3:1 (insert:vector) ratio 4:1 (insert:vector) ratio
D112 (insert) 0 µL 1,4 µL 1,9 µL
D187 (vector) 2,5 µL 2,5 µL 2,5 µL
T4 DNA ligase 10X buffer 2 µL 2 µL 2 µL
T4 DNA ligase 1 µL 1 µL 1 µL
water 14,5 µL 13,1 µL 12,6 µL
  • overnight reaction at about 16°C.


Construction of rbs-FlhDC-mRFP-LVA-tripart (L172)

Part icon rbs.pngIcon coding.pngPart icon reporter.pngPart icon terminator.pngPart icon terminator.png

Ligation L172

Digestion n° Name Digested by DNA concentration
D186 rbs-FlhDC EcoRI & SpeI (insert) ~24 µg/mL
D187 mRFP-LVA-tripart EcoRI & XbaI (vector) 16 µg/mL


Reaction mixtures (3:1 (insert:vector) ratio)

Control Sample
D186 (insert) 0 µL 1 µL
D187 (vector) 2,5 µL 2,5 µL
T4 DNA ligase 10X buffer 2 µL 2 µL
T4 DNA ligase 1 µL 1 µL
water 14,5 µL 13,5 µL
  • overnight reaction at about 16°C.
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