Team:Paris/July 24

From 2008.igem.org

(Difference between revisions)
(→MiniPreps)
(→MiniPreps)
 
(9 intermediate revisions not shown)
Line 4: Line 4:
== MiniPreps ==
== MiniPreps ==
-
*The protocol of MiniPrep of Promega has been done on all the clones cultivated on the 23th.
+
*The Promega MiniPreps protocol has been used on all the clones cultivated on the 23th.
{| border="1"  
{| border="1"  
Line 10: Line 10:
|align="center"|'''Biobrick'''
|align="center"|'''Biobrick'''
|align="center"|'''Description'''
|align="center"|'''Description'''
-
|-
 
-
|align="center"|MP100
 
-
|align="center"|[http://partsregistry.org/Part:BBa_B0034 B0034]
 
-
|align="center"|Strongest RBS (Efficiency = 1)
 
-
|-
 
-
|align="center"|MP101
 
-
|align="center"|[http://partsregistry.org/Part:BBa_J23101 J23101]
 
-
|align="center"|Strongest Constitutive promoter
 
-
|-
 
-
|align="center"|MP102
 
-
|align="center"|[http://partsregistry.org/Part:BBa_J23109 J23109]
 
-
|align="center"|Weak Constitutive promoter
 
-
|-
 
-
|align="center"|MP103
 
-
|align="center"|[http://partsregistry.org/Part:BBa_R0079 R0079]
 
-
|align="center"|promoter pLas (lasr & PAI regulated)
 
-
|-
 
-
|align="center"|MP104
 
-
|align="center"|[http://partsregistry.org/Part:BBa_R0040 R0040]
 
-
|align="center"|tetR repressible promoter
 
-
|-
 
-
|align="center"|MP105
 
-
|align="center"|[http://partsregistry.org/Part:BBa_S03154 S03154]
 
-
|align="center"|B0034(rbs) ->LasI
 
-
|-
 
-
|align="center"|MP106
 
-
|align="center"|[http://partsregistry.org/Part:BBa_S03879 S03879]
 
-
|align="center"|B0034(rbs) -> TetR
 
-
|-
 
-
|align="center"|MP107
 
-
|align="center"|[http://partsregistry.org/Part:BBa_C0079 C0079]
 
-
|align="center"|lasR activator with LVA Tag
 
-
|-
 
-
|align="center"|MP108
 
-
|align="center"|[http://partsregistry.org/Part:BBa_C0179 C0179]
 
-
|align="center"|lasR activator
 
-
|-
 
-
|align="center"|MP109
 
-
|align="center"|[http://partsregistry.org/Part:BBa_E0030 E0030]
 
-
|align="center"|YFP
 
-
|-
 
-
|align="center"|MP110
 
-
|align="center"|[http://partsregistry.org/Part:BBa_E0040 E0040]
 
-
|align="center"|GFP
 
-
|-
 
-
|align="center"|MP111
 
-
|align="center"|[http://partsregistry.org/Part:BBa_E1010 E1010]
 
-
|align="center"|mRFP
 
|-
|-
|align="center"|MP116
|align="center"|MP116
Line 90: Line 42:
== Digestion ==
== Digestion ==
-
 
===Digestion Mix===
===Digestion Mix===
-
10µl of Miniprep (clone 1 or 2)<br>
+
10µl of Miniprep (26 aug.) <br>
12.5µl of water<br>
12.5µl of water<br>
2.5µl of Buffer N°2<br>
2.5µl of Buffer N°2<br>
Line 101: Line 52:
1µl of enzyme 1<br>
1µl of enzyme 1<br>
1µl of enzyme 2<br>
1µl of enzyme 2<br>
 +
 +
* Incubation 1h at 37°C with the first enzyme
 +
* Add the second enzyme
 +
* Incubation 1h at 37°C with the second enzyme
 +
* Store on ice
 +
* Revelation of the digestion by electrophoresis on agarose gel
Line 106: Line 63:
=== Results of digestions : Electrophoresis ===
=== Results of digestions : Electrophoresis ===
 +
conditions :
 +
* 10µl of ladder 1 kb (except for gel n°7 : 100 pb)
 +
* 30µl of digestion added with 5µl of loading Dye 6x
 +
* migration ~30min at 100W
 +
* Gel 1, 2, 3, 4, 5, 6, 8 = '''0.8%'''   
 +
* Gel 7 = '''1,2%'''
-
gel1 [[Image:| gel1|200px]]
 
-
gel2 [[Image:| gel2|200px]]
 
-
gel3 [[Image:| gel3|200px]]
 
-
gel4 [[Image:| gel4|200px]]<br>
 
 +
gel1 [[Image:Gel1.jpg| gel1|200px]]
 +
gel2 [[Image:Gel2.jpg| gel2|200px]]
 +
gel3 [[Image:Gel3.jpg| gel3|200px]]
 +
gel4 [[Image:Gel4.jpg| gel4|200px]]<br>
-
gel5 [[Image:‎| gel5|200px]]
+
 
-
gel6 [[Image:‎| gel6|200px]]
+
gel5 [[Image:Gel_5.jpg‎| gel5|200px]]
-
gel7 [[Image:| gel7|100px]]
+
gel6 [[Image:Gel_6.jpg‎| gel6|200px]]
-
gel8 [[Image:|100px]]
+
gel7 [[Image:Gel_7.jpg‎| gel7|100px]]
 +
gel8 [[Image:Gel_8.jpg‎| gel8|100px]]
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! rowspan="2"| 2057 pb
! rowspan="2"| 2057 pb
! rowspan="2"| 34 pb
! rowspan="2"| 34 pb
-
! rowspan="2"|  
+
! rowspan="2"| not digested
-
! rowspan="2"|  
+
! rowspan="2"| not digested
-
! rowspan="2"|  
+
! rowspan="2"| 8
-
! rowspan="2"|  
+
! rowspan="2"| 5-6
|-
|-
|align=center|2
|align=center|2
Line 156: Line 120:
|align=center| 2076 pb
|align=center| 2076 pb
|align=center| 15 pb
|align=center| 15 pb
-
|align=center|
+
|align=center| 2100 pb
-
|align=center|  
+
|align=center| -
-
|align=center|  
+
|align=center| 1
-
|align=center|  
+
|align=center| 2
|-
|-
|align=center|4
|align=center|4
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! rowspan="2"| 2077 pb
! rowspan="2"| 2077 pb
! rowspan="2"| 14 pb
! rowspan="2"| 14 pb
-
! rowspan="2"|  
+
! rowspan="2"| 2000 pb
-
! rowspan="2"|  
+
! rowspan="2"| -
-
|align=center|  
+
|align=center| 1
-
|align=center|  
+
|align=center| 3-4
|-
|-
|align=center|5
|align=center|5
-
|align=center|  
+
|align=center| 1
-
|align=center|  
+
|align=center| 5-6
|-
|-
! rowspan="2" style="background: #ccccff;" | D101
! rowspan="2" style="background: #ccccff;" | D101
Line 184: Line 148:
! rowspan="2"| 2100 pb
! rowspan="2"| 2100 pb
! rowspan="2"| 883 pb
! rowspan="2"| 883 pb
-
! rowspan="2"|  
+
! rowspan="2"| 2000 pb
-
! rowspan="2"|  
+
! rowspan="2"| 900 pb
-
|align=center|  
+
|align=center| 1
-
|align=center|  
+
|align=center| 7-8
|-
|-
|align=center|2
|align=center|2
-
|align=center|  
+
|align=center| 1
-
|align=center|  
+
|align=center| 9-10
|-
|-
! rowspan="1" style="background: #ccccff;" | D102
! rowspan="1" style="background: #ccccff;" | D102
Line 201: Line 165:
! rowspan="1"| 2100 pb
! rowspan="1"| 2100 pb
! rowspan="1"| 883 pb
! rowspan="1"| 883 pb
-
|align=center|  
+
|align=center| 2000 pb
-
|align=center|  
+
|align=center| 900 pb
-
|align=center|  
+
|align=center| 1 & 2
-
|align=center|  
+
|align=center| 11-12 & 2-3
|-
|-
! rowspan="2" style="background: #ccccff;" | D103
! rowspan="2" style="background: #ccccff;" | D103
Line 214: Line 178:
! rowspan="2"| 2222 pb
! rowspan="2"| 2222 pb
! rowspan="2"| 14 pb
! rowspan="2"| 14 pb
-
! rowspan="2"|  
+
! rowspan="2"| 2500 pb
-
! rowspan="2"|  
+
! rowspan="2"| -
-
|align=center|  
+
|align=center| 2
-
|align=center|  
+
|align=center| 4-5
|-
|-
|align=center|2
|align=center|2
-
|align=center|  
+
|align=center| 2
-
|align=center|  
+
|align=center| 6-7
|-
|-
! rowspan="2" style="background: #ccccff;" | D104
! rowspan="2" style="background: #ccccff;" | D104
Line 231: Line 195:
! rowspan="2"| 2119 pb
! rowspan="2"| 2119 pb
! rowspan="2"| 14 pb
! rowspan="2"| 14 pb
-
! rowspan="2"|  
+
! rowspan="2"| 2500 pb
-
! rowspan="2"|  
+
! rowspan="2"| -
-
|align=center|  
+
|align=center| 2
-
|align=center|  
+
|align=center| 8-9
|-
|-
|align=center|2
|align=center|2
-
|align=center|  
+
|align=center| 2
-
|align=center|  
+
|align=center| 10-11
|-
|-
! rowspan="4" style="background: #ccccff;" | D105
! rowspan="4" style="background: #ccccff;" | D105
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! rowspan="1"| 2750 pb
! rowspan="1"| 2750 pb
! rowspan="1"| 14 pb
! rowspan="1"| 14 pb
-
|align=center|  
+
|align=center| 3000 pb
-
|align=center|  
+
|align=center| -
-
|align=center|  
+
|align=center| 2 & 3
-
|align=center|  
+
|align=center| 12 & 2
|-
|-
|align=center|2
|align=center|2
Line 259: Line 223:
! rowspan="2"| 2057 pb
! rowspan="2"| 2057 pb
! rowspan="2"| 707 pb
! rowspan="2"| 707 pb
-
! rowspan="2"|
+
! rowspan="2"| 2000 pb
-
! rowspan="2"|  
+
! rowspan="2"| 700 pb
-
|align=center|  
+
|align=center| 3
-
|align=center|  
+
|align=center| 3-4
|-
|-
|align=center|3
|align=center|3
-
|align=center|  
+
|align=center| 3
-
|align=center|  
+
|align=center| 5-6
|-
|-
|align=center|4
|align=center|4
Line 274: Line 238:
| align=center| 2056 pb
| align=center| 2056 pb
| align=center| 708 pb
| align=center| 708 pb
-
|align=center|  
+
|align=center| 2000 pb
-
|align=center|  
+
|align=center| 700 pb
-
|align=center|  
+
|align=center| 3
-
|align=center|  
+
|align=center| 7-8
|-
|-
! rowspan="4" style="background: #ccccff;" | D106
! rowspan="4" style="background: #ccccff;" | D106
Line 287: Line 251:
| align=center| 2768 pb
| align=center| 2768 pb
| align=center|14 pb
| align=center|14 pb
-
|align=center|  
+
|align=center| 3000 pb
-
|align=center|  
+
|align=center| -
-
|align=center|  
+
|align=center| 3
-
|align=center|  
+
|align=center| 9-10
|-
|-
|align=center|2
|align=center|2
Line 298: Line 262:
! rowspan="2"| 2057 pb
! rowspan="2"| 2057 pb
! rowspan="2"| 725 pb
! rowspan="2"| 725 pb
-
! rowspan="2"|  
+
! rowspan="2"| 2000 pb
-
! rowspan="2"|  
+
! rowspan="2"| 700 pb
-
|align=center|  
+
|align=center| 3
-
|align=center|  
+
|align=center| 11-12
|-
|-
|align=center|3
|align=center|3
-
|align=center|  
+
|align=center| 4
-
|align=center|  
+
|align=center| 2-3
|-
|-
|align=center|4
|align=center|4
Line 313: Line 277:
| align=center| 2756 pb
| align=center| 2756 pb
| align=center| 726 pb
| align=center| 726 pb
-
|align=center|  
+
|align=center| 2500 pb
-
|align=center|  
+
|align=center| 700 pb
-
|align=center|  
+
|align=center| 4
-
|align=center|  
+
|align=center| 4-5
|-
|-
! rowspan="2" style="background: #ccccff;" |D107
! rowspan="2" style="background: #ccccff;" |D107
Line 326: Line 290:
|align=center| 4402 pb
|align=center| 4402 pb
|align=center| 779 pb
|align=center| 779 pb
-
|align=center|  
+
|align=center| 6000 pb
-
|align=center|  
+
|align=center| 1000 pb
-
|align=center|  
+
|align=center| 4
-
|align=center|  
+
|align=center| 6-7
|-
|-
|align=center| 2
|align=center| 2
Line 337: Line 301:
|align=center| 4003 pb
|align=center| 4003 pb
|align=center| 778 pb
|align=center| 778 pb
-
|align=center|  
+
|align=center| 5000 pb
-
|align=center|  
+
|align=center| 100 pb
-
|align=center|  
+
|align=center| 6
-
|align=center|  
+
|align=center| 9
|-
|-
! rowspan="2" style="background: #ccccff;"|D108
! rowspan="2" style="background: #ccccff;"|D108
Line 350: Line 314:
|align=center| 4402 pb
|align=center| 4402 pb
|align=center| 746 pb
|align=center| 746 pb
-
|align=center|
+
|align=center| 2500 pb
-
|align=center|  
+
|align=center| 1000 pb
-
|align=center|  
+
|align=center| 4
-
|align=center|  
+
|align=center| 8-9
|-
|-
|align=center| 2
|align=center| 2
Line 361: Line 325:
|align=center| 4403 pb
|align=center| 4403 pb
|align=center| 745 pb
|align=center| 745 pb
-
|align=center|  
+
|align=center| 2500 pb
-
|align=center|  
+
|align=center| 1000 pb
-
|align=center|  
+
|align=center| 6
-
|align=center|  
+
|align=center| 10
|-
|-
! rowspan="2" style="background: #ccccff;"|D109
! rowspan="2" style="background: #ccccff;"|D109
Line 374: Line 338:
|align=center| 3166 pb
|align=center| 3166 pb
|align=center| 746 pb
|align=center| 746 pb
-
|align=center|  
+
|align=center| 4000 pb
-
|align=center|  
+
|align=center| 1000 pb
-
|align=center|  
+
|align=center| 4
-
|align=center|  
+
|align=center| 10-11
|-
|-
|align=center| 2
|align=center| 2
Line 385: Line 349:
|align=center| 3167 pb
|align=center| 3167 pb
|align=center| 745 pb
|align=center| 745 pb
-
|align=center|  
+
|align=center| 3000 pb
-
|align=center|  
+
|align=center| 1000 pb
-
|align=center|  
+
|align=center| 6
-
|align=center|  
+
|align=center| 11
|-
|-
! rowspan="2" style="background: #ccccff;"|D110
! rowspan="2" style="background: #ccccff;"|D110
Line 398: Line 362:
|align=center| 2056 pb
|align=center| 2056 pb
|align=center| 743 pb
|align=center| 743 pb
-
|align=center|  
+
|align=center| 2000 pb
-
|align=center|  
+
|align=center| 800 pb
-
|align=center|  
+
|align=center| 4 & 5
-
|align=center|  
+
|align=center| 12 & 2
|-
|-
|align=center| 2
|align=center| 2
Line 409: Line 373:
|align=center| 2057 pb
|align=center| 2057 pb
|align=center| 742 pb
|align=center| 742 pb
-
|align=center|  
+
|align=center| 2500 pb
-
|align=center|  
+
|align=center| 1000 pb
-
|align=center|  
+
|align=center| 6
-
|align=center|  
+
|align=center| 12
|-
|-
! rowspan="2" style="background: #ccccff;"|D111
! rowspan="2" style="background: #ccccff;"|D111
Line 422: Line 386:
|align=center| 4402 pb
|align=center| 4402 pb
|align=center| 704 pb
|align=center| 704 pb
-
|align=center|  
+
|align=center| 4000 pb
-
|align=center|  
+
|align=center| 600 pb
-
|align=center|  
+
|align=center| 5
-
|align=center|  
+
|align=center| 3-4
|-
|-
|align=center| 2
|align=center| 2
Line 433: Line 397:
|align=center| 4403 pb
|align=center| 4403 pb
|align=center| 703 pb
|align=center| 703 pb
-
|align=center|  
+
|align=center| 4500 pb
-
|align=center|  
+
|align=center| 700 pb
-
|align=center|  
+
|align=center| 8
-
|align=center|  
+
|align=center| 2-3
|-
|-
-
! rowspan="2" style="background: #ccccff;"|D116
+
! rowspan="1" style="background: #ccccff;"|D116
-
! rowspan="2" style="background: #ccccff;"|J23100
+
! rowspan="1" style="background: #ccccff;"|J23100
|align=center|1
|align=center|1
-
! rowspan="2"| SpeI
+
! rowspan="1"| SpeI
-
! rowspan="2"| PstI
+
! rowspan="1"| PstI
-
! rowspan="2"| BV
+
! rowspan="1"| BV
-
! rowspan="2"| 2100 pb
+
! rowspan="1"| 2100 pb
-
! rowspan="2"| 883 pb
+
! rowspan="1"| 883 pb
-
! colspan="4"|  
+
! colspan="4"| Not realised
|align=center|
|align=center|
|-
|-
-
|align=center|2
+
! rowspan="1" style="background: #ccccff;"|D117
-
|
+
! rowspan="1" style="background: #ccccff;"|J23107
-
|
+
-
|-
+
-
! rowspan="2" style="background: #ccccff;"|D117
+
-
! rowspan="2" style="background: #ccccff;"|J23107
+
|align=center|1
|align=center|1
-
! rowspan="2"| SpeI
+
! rowspan="1"| SpeI
-
! rowspan="2"| PstI
+
! rowspan="1"| PstI
-
! rowspan="2"| BV
+
! rowspan="1"| BV
-
! rowspan="2"| 2100 pb
+
! rowspan="1"| 2100 pb
-
! rowspan="2"| 883 pb
+
! rowspan="1"| 883 pb
-
|align=center|  
+
|align=center| 2000 pb
-
|align=center|
+
|align=center| 800 pb
-
|align=center|
+
|align=center| 5
-
|align=center|
+
|align=center| 5-6
-
|align=center|
+
|-
|-
-
|align=center|2
+
! rowspan="1" style="background: #ccccff;"|D118
-
|
+
! rowspan="1" style="background: #ccccff;"|B0015
-
|-
+
-
! rowspan="2" style="background: #ccccff;"|D118
+
-
! rowspan="2" style="background: #ccccff;"|B0015
+
|align=center|1
|align=center|1
|align=center|EcoRI
|align=center|EcoRI
|align=center|XbaI
|align=center|XbaI
-
! rowspan="2"| FV
+
! rowspan="1"| FV
-
! rowspan="2"| 3303 pb
+
! rowspan="1"| 3303 pb
-
! rowspan="2"| 15 pb
+
! rowspan="1"| 15 pb
-
|align=center|  
+
|align=center| 3000 pb
-
|align=center|  
+
|align=center| -
-
|align=center|  
+
|align=center| 5
-
|align=center|  
+
|align=center| 7-8
-
|align=center|
+
-
|-
+
-
|align=center|2
+
-
|
+
|-
|-
-
! rowspan="2" style="background: #ccccff;"|D119
+
! rowspan="1" style="background: #ccccff;"|D119
-
! rowspan="2" style="background: #ccccff;"|I0500
+
! rowspan="1" style="background: #ccccff;"|I0500
|align=center|1
|align=center|1
|align=center|SpeI
|align=center|SpeI
|align=center|PstI
|align=center|PstI
-
! rowspan="2"| FV
+
! rowspan="1"| FV
-
! rowspan="2"| 5621 pb
+
! rowspan="1"| 5621 pb
-
! rowspan="2"| 14 pb
+
! rowspan="1"| 14 pb
-
! rowspan="2"| 
+
|align=center| 6000 - 3000 pb
-
|align=center|
+
|align=center| -
-
|align=center|
+
|align=center| 5
-
|align=center|
+
|align=center| 9-10
-
|align=center|
+
-
|-
+
-
|align=center|2
+
-
|
+
-
|
+
|-
|-
! rowspan="5" style="background: #ccccff;" | D120
! rowspan="5" style="background: #ccccff;" | D120
Line 513: Line 460:
! rowspan="2"| 2057 pb
! rowspan="2"| 2057 pb
! rowspan="2"| 37 pb
! rowspan="2"| 37 pb
-
! rowspan="2"|
+
! rowspan="2"| not digested
-
! rowspan="2"|
+
! rowspan="2"| not digested
-
|align=center|
+
|align=center| 7
-
|align=center|  
+
|align=center| 2
|-
|-
|align=center|2
|align=center|2
-
|align=center|  
+
|align=center| 7
-
|align=center|  
+
|align=center| 5-6
|-
|-
|align=center|3
|align=center|3
Line 528: Line 475:
|align=center| 2079 pb
|align=center| 2079 pb
|align=center| 15 pb
|align=center| 15 pb
-
|align=center|  
+
|align=center| 1600 pb
-
|align=center|  
+
|align=center| -
-
|align=center|  
+
|align=center| 6
-
|align=center|  
+
|align=center| 2
|-
|-
|align=center|4
|align=center|4
Line 539: Line 486:
! rowspan="2"| 2080 pb
! rowspan="2"| 2080 pb
! rowspan="2"| 14 pb
! rowspan="2"| 14 pb
-
! rowspan="2"|  
+
! rowspan="2"| 3000 - 2000 pb
-
! rowspan="2"|  
+
! rowspan="2"| -
-
|align=center|  
+
|align=center| 5
-
|align=center|  
+
|align=center| 11
|-
|-
|align=center|5
|align=center|5
-
|align=center|  
+
|align=center| 5
-
|align=center|  
+
|align=center| 12
|-
|-
! rowspan="1" style="background: #ccccff;"|D121
! rowspan="1" style="background: #ccccff;"|D121
Line 553: Line 500:
|align=center|XbaI
|align=center|XbaI
|align=center|PstI
|align=center|PstI
-
|align=center| BI
+
|align=center| FV
|align=center| 2057 pb
|align=center| 2057 pb
|align=center| 939 pb
|align=center| 939 pb
-
|align=center|  
+
|align=center| 3000 pb
-
|align=center|  
+
|align=center| Not digested
-
|align=center|  
+
|align=center| 6
-
|align=center|  
+
|align=center| 3-5
|-
|-
! rowspan="1" style="background: #ccccff;"|D122
! rowspan="1" style="background: #ccccff;"|D122
Line 566: Line 513:
|align=center|XbaI
|align=center|XbaI
|align=center|PstI
|align=center|PstI
-
|align=center| BI
+
|align=center| FV
|align=center| 2057 pb
|align=center| 2057 pb
|align=center| 900 pb
|align=center| 900 pb
-
|align=center|  
+
|align=center| 2500 pb
-
|align=center|  
+
|align=center| Not digested
-
|align=center|  
+
|align=center| 6
-
|align=center|  
+
|align=center| 6-8
|}
|}
 +
 +
 +
==> Conclusion: Most of the digestion have succeed
 +
 +
== Extraction of the DNA  ==
 +
 +
 +
* Cutting of the parts of interest, for all the digestion that have migrated on the gels
 +
* Store of all the piece of gel O/N at -20°C.

Latest revision as of 14:39, 31 July 2008

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Contents

MiniPreps

  • The Promega MiniPreps protocol has been used on all the clones cultivated on the 23th.
Name Biobrick Description
MP116 [http://partsregistry.org/Part:BBa_J23100 J23100] Strong constitutive promoter in J61002
MP117 [http://partsregistry.org/Part:BBa_J23107 J23107] Medium constitutive promoter in J61002
MP118 [http://partsregistry.org/Part:BBa_B0015 B0015] Double terminator
MP119 [http://partsregistry.org/Part:BBa_I0500 I0500] AraC pBAD
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

Digestion

Digestion Mix

10µl of Miniprep (26 aug.)
12.5µl of water
2.5µl of Buffer N°2
0.25µl of BSA 100x
1µl of enzyme 1
1µl of enzyme 2

  • Incubation 1h at 37°C with the first enzyme
  • Add the second enzyme
  • Incubation 1h at 37°C with the second enzyme
  • Store on ice
  • Revelation of the digestion by electrophoresis on agarose gel


Results of digestions : Electrophoresis

conditions :

  • 10µl of ladder 1 kb (except for gel n°7 : 100 pb)
  • 30µl of digestion added with 5µl of loading Dye 6x
  • migration ~30min at 100W
  • Gel 1, 2, 3, 4, 5, 6, 8 = 0.8%
  • Gel 7 = 1,2%


gel1 gel1 gel2 gel2 gel3 gel3 gel4 gel4


gel5 gel5 gel6 gel6 gel7 gel7 gel8 gel8


Name BioBrick Tube N° Enz 1 Enz 2 Obs Exp Size Matrix Exp Size BB Mea Size Matrix Mea Size BB Gel Band
D100 B0034 1 XbaI PstI BI 2057 pb 34 pb not digested not digested 8 5-6
2
3 EcoRI XbaI FV 2076 pb 15 pb 2100 pb - 1 2
4 SpeI PstI BV 2077 pb 14 pb 2000 pb - 1 3-4
5 1 5-6
D101 J23101 1 SpeI PstI BV 2100 pb 883 pb 2000 pb 900 pb 1 7-8
2 1 9-10
D102 J23109 1 SpeI PstI BV 2100 pb 883 pb 2000 pb 900 pb 1 & 2 11-12 & 2-3
D103 R0079 1 SpeI PstI BV 2222 pb 14 pb 2500 pb - 2 4-5
2 2 6-7
D104 R0040 1 SpeI PstI BV 2119 pb 14 pb 2500 pb - 2 8-9
2 2 10-11
D105 S03154 1 SpeI PstI BV 2750 pb 14 pb 3000 pb - 2 & 3 12 & 2
2 XbaI PstI BI 2057 pb 707 pb 2000 pb 700 pb 3 3-4
3 3 5-6
4 EcoRI SpeI FI 2056 pb 708 pb 2000 pb 700 pb 3 7-8
D106 S03879 1 SpeI PstI BV 2768 pb 14 pb 3000 pb - 3 9-10
2 XbaI PstI BI 2057 pb 725 pb 2000 pb 700 pb 3 11-12
3 4 2-3
4 EcoRI SpeI FI 2756 pb 726 pb 2500 pb 700 pb 4 4-5
D107 C0079 1 EcoRI SpeI FI 4402 pb 779 pb 6000 pb 1000 pb 4 6-7
2 XbaI PstI BI 4003 pb 778 pb 5000 pb 100 pb 6 9
D108 C0179 1 EcoRI SpeI FI 4402 pb 746 pb 2500 pb 1000 pb 4 8-9
2 XbaI PstI BI 4403 pb 745 pb 2500 pb 1000 pb 6 10
D109 E0030 1 EcoRI SpeI FI 3166 pb 746 pb 4000 pb 1000 pb 4 10-11
2 XbaI PstI BI 3167 pb 745 pb 3000 pb 1000 pb 6 11
D110 E0040 1 EcoRI SpeI FI 2056 pb 743 pb 2000 pb 800 pb 4 & 5 12 & 2
2 XbaI PstI BI 2057 pb 742 pb 2500 pb 1000 pb 6 12
D111 E1010 1 EcoRI SpeI FI 4402 pb 704 pb 4000 pb 600 pb 5 3-4
2 XbaI PstI BI 4403 pb 703 pb 4500 pb 700 pb 8 2-3
D116 J23100 1 SpeI PstI BV 2100 pb 883 pb Not realised
D117 J23107 1 SpeI PstI BV 2100 pb 883 pb 2000 pb 800 pb 5 5-6
D118 B0015 1 EcoRI XbaI FV 3303 pb 15 pb 3000 pb - 5 7-8
D119 I0500 1 SpeI PstI FV 5621 pb 14 pb 6000 - 3000 pb - 5 9-10
D120 B0030 1 XbaI PstI BI 2057 pb 37 pb not digested not digested 7 2
2 7 5-6
3 EcoRI XbaI FV 2079 pb 15 pb 1600 pb - 6 2
4 SpeI PstI BV 2080 pb 14 pb 3000 - 2000 pb - 5 11
5 5 12
D121 E0422 1 XbaI PstI FV 2057 pb 939 pb 3000 pb Not digested 6 3-5
D122 E0840 1 XbaI PstI FV 2057 pb 900 pb 2500 pb Not digested 6 6-8


==> Conclusion: Most of the digestion have succeed

Extraction of the DNA

  • Cutting of the parts of interest, for all the digestion that have migrated on the gels
  • Store of all the piece of gel O/N at -20°C.
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