Team:Paris/July 24

From 2008.igem.org

(Difference between revisions)
(→Results of digestions : Electrophoresis)
(→MiniPreps)
 
(10 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 12: Line 12:
|-
|-
|align="center"|MP116
|align="center"|MP116
-
|align="center"|J23100
+
|align="center"|[http://partsregistry.org/Part:BBa_J23100 J23100]
-
|align="center"|
+
|align="center"|Strong constitutive promoter in J61002
|-
|-
|align="center"|MP117
|align="center"|MP117
-
|align="center"|J23107
+
|align="center"|[http://partsregistry.org/Part:BBa_J23107 J23107]
-
|align="center"|
+
|align="center"|Medium constitutive promoter in J61002
|-
|-
|align="center"|MP118
|align="center"|MP118
-
|align="center"|B0015
+
|align="center"|[http://partsregistry.org/Part:BBa_B0015 B0015]
-
|align="center"|
+
|align="center"|Double terminator
|-
|-
|align="center"|MP119
|align="center"|MP119
-
|align="center"|I0500
+
|align="center"|[http://partsregistry.org/Part:BBa_I0500 I0500]
-
|align="center"|
+
|align="center"|AraC pBAD
|-
|-
|align="center"|MP120
|align="center"|MP120
-
|align="center"|B0030
+
|align="center"|[http://partsregistry.org/Part:BBa_B0030 B0030]
-
|align="center"|
+
|align="center"|Strong RBS (Efficiency = 0,6)
|-
|-
|align="center"|MP121
|align="center"|MP121
-
|align="center"|E0422
+
|align="center"|[http://partsregistry.org/Part:BBa_E0422 E0422]
-
|align="center"|
+
|align="center"|ECFP (RBS+LVA+Term)
|-
|-
-
|align="center"|MP122
+
|align="center"| MP122
-
|align="center"|E0840
+
|align="center"|[http://partsregistry.org/Part:BBa_E0840 E0840]
-
|align="center"|
+
|align="center"|gfp tri-part; strong rbs
 +
|
|}
|}
== 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 52: 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
=== 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]]
+
gel1 [[Image:Gel1.jpg| gel1|200px]]
-
gel2 [[Image:| gel2|200px]]
+
gel2 [[Image:Gel2.jpg| gel2|200px]]
-
gel3 [[Image:| gel3|200px]]
+
gel3 [[Image:Gel3.jpg| gel3|200px]]
-
gel4 [[Image:| gel4|200px]]<br>
+
gel4 [[Image:Gel4.jpg| gel4|200px]]<br>
-
gel5 [[Image:‎| gel5|200px]]
+
gel5 [[Image:Gel_5.jpg‎| gel5|200px]]
-
gel6 [[Image:‎| gel6|200px]]
+
gel6 [[Image:Gel_6.jpg‎| gel6|200px]]
-
gel7 [[Image:| gel7|100px]]
+
gel7 [[Image:Gel_7.jpg‎| gel7|100px]]
-
gel8 [[Image:|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 107: 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
Line 118: Line 131:
! 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 135: 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 152: 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 165: 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 182: 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
Line 199: Line 212:
! 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 210: 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 225: 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 238: 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 249: 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 264: 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 277: 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 288: 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 301: 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 312: 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 325: 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 336: 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 349: 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 360: 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 373: 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 384: 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="2" style="background: #ccccff;"|D118
+
! rowspan="1" style="background: #ccccff;"|D118
-
! rowspan="2" style="background: #ccccff;"|B0015
+
! rowspan="1" 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 464: 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 479: 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 490: 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 504: 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 517: 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 J23100 Strong constitutive promoter in J61002
MP117 J23107 Medium constitutive promoter in J61002
MP118 B0015 Double terminator
MP119 I0500 AraC pBAD
MP120 B0030 Strong RBS (Efficiency = 0,6)
MP121 E0422 ECFP (RBS+LVA+Term)
MP122 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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