Team:Paris/July 26

From 2008.igem.org

(Difference between revisions)
(→MiniPreps)
(→Extraction of the DNA)
 
(22 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 25th.
+
 
 +
* Use of Promega's protocol on all the clones cultivated on the 25th.
 +
* Preparation of 50µl of minipreps in double.
 +
 
{| border="1"  
{| border="1"  
Line 89: Line 92:
|}
|}
-
== 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 102: Line 106:
1µl of enzyme 2<br>
1µl of enzyme 2<br>
-
===list of digestion===
+
* 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
-
{| border="1"  
+
 
-
|align="center"|'''Name'''
+
 
-
|align="center"|'''Miniprep'''
+
=== Results of digestions : Electrophoresis ===
-
|align="center"|'''Biobrick'''
+
 
-
|align="center"|'''Enzyme 1'''
+
'''conditions :'''
-
|align="center"|'''Enzyme 2'''
+
-
|align="center"|'''Nber tube'''
+
* 10µl of ladder 1 kb (except for gel n°6 : 100 pb)
 +
* 30µl of digestion added with 5µl of loading Dye 6x
 +
* migration ~30min at 100W
 +
* Gel 1, 2, 3, 4, 5 = '''0.8%'''   
 +
* Gel 6 = '''1,5%'''
 +
 
 +
 
 +
gel1 [[Image:080726gel_1.jpg| gel1|200px]]
 +
gel2 [[Image:080726gel_2.jpg| gel2|200px]]
 +
gel3 [[Image:080726gel_3.jpg| gel3|200px]]
 +
gel4 [[Image:080726gel_4.jpg| gel4|200px]]<br>
 +
 
 +
 
 +
gel5 [[Image:080726gel_5.jpg| gel5|100px]]
 +
gel6 [[Image:080726gel_6.jpg| gel6|100px]]
 +
 
 +
 
 +
 
 +
{|- border="1"
 +
|align=center style="background: #649CD7;"|'''Name'''
 +
|align=center style="background: #649CD7;"|'''BioBrick'''
 +
|align=center style="background: #649CD7;"|'''Tube N°'''
 +
|align=center style="background: #649CD7;"|'''Enz 1'''
 +
|align=center style="background: #649CD7;"|'''Enz 2'''
 +
|align=center style="background: #649CD7;"|'''Obs'''
 +
|align=center style="background: #649CD7;"|'''Exp Size Matrix'''
 +
|align=center style="background: #649CD7;"|'''Exp Size BB'''
 +
|align=center style="background: #649CD7;"|'''Mea Size Matrix'''
 +
|align=center style="background: #649CD7;"|'''Mea Size BB'''
 +
|align=center style="background: #649CD7;"|'''Gel'''
 +
|align=center style="background: #649CD7;"|'''Band'''
|-
|-
-
|align="center"|D100
+
! rowspan="2" style="background: #D4E2EF;" | D100
-
|align="center"|MP100
+
! rowspan="2" style="background: #D4E2EF;" | B0034
-
|align="center"|B0034
+
|align=center|1
-
|align="center"|XbaI
+
! rowspan="2"| XbaI
-
|align="center"|PstI
+
! rowspan="2"| PstI
-
|align="center"|2
+
! rowspan="2"| BI
 +
! rowspan="2"| 2057 pb
 +
! rowspan="2"| 34 pb
 +
! rowspan="2"| 1800 pb
 +
! rowspan="2"| 40 pb
 +
! rowspan="1"| 6
 +
! rowspan="1"| 2
|-
|-
-
|align="center"|
+
|align=center|2
-
|align="center"|
+
! rowspan="1"| 6
-
|align="center"|
+
! rowspan="1"| 3
-
|align="center"|EcoRI
+
-
|align="center"|XbaI
+
-
|align="center"|1
+
|-
|-
-
|align="center"|
+
! rowspan="1" style="background: #D4E2EF;" | D101
-
|align="center"|
+
! rowspan="1" style="background: #D4E2EF;" | B0034
-
|align="center"|
+
|align=center|3
-
|align="center"|SpeI
+
|align=center| EcoRI
-
|align="center"|PstI
+
|align=center| XbaI
-
|align="center"|2
+
|align=center| FV
 +
|align=center| 2076 pb
 +
|align=center| 15 pb
 +
|align=center| 2000 pb
 +
|align=center| -
 +
|align=center| 1
 +
|align=center| 2
|-
|-
-
|align="center"|D101
+
! rowspan="2" style="background: #D4E2EF;" | D102
-
|align="center"|MP101
+
! rowspan="2" style="background: #D4E2EF;" | B0034
-
|align="center"|J23101
+
|align=center|4
-
|align="center"|SpeI
+
! rowspan="2"| SpeI
-
|align="center"|PstI
+
! rowspan="2"| PstI
-
|align="center"|2
+
! rowspan="2"| BV
 +
! rowspan="2"| 2077 pb
 +
! rowspan="2"| 14 pb
 +
! rowspan="2"| 2000 pb
 +
! rowspan="2"| -
 +
|align=center| 1
 +
|align=center| 3
|-
|-
-
|align="center"|D102
+
|align=center|5
-
|align="center"|MP102
+
|align=center| 1
-
|align="center"|J23109
+
|align=center| 4
-
|align="center"|SpeI
+
-
|align="center"|PstI
+
-
|align="center"|2
+
|-
|-
-
|align="center"|D103
+
! rowspan="2" style="background: #D4E2EF;" | D103
-
|align="center"|MP103
+
! rowspan="2" style="background: #D4E2EF;" | J23101
-
|align="center"|R0079
+
|align=center|1
-
|align="center"|SpeI
+
! rowspan="2"| SpeI
-
|align="center"|PstI
+
! rowspan="2"| PstI
-
|align="center"|2
+
! rowspan="2"| BV
 +
! rowspan="2"| 2100 pb
 +
! rowspan="2"| 883 pb
 +
! rowspan="2"| 2000 pb
 +
! rowspan="2"| 700 pb
 +
|align=center| 1
 +
|align=center| 5
|-
|-
-
|align="center"|D104
+
|align=center|2
-
|align="center"|MP104
+
|align=center| 1
-
|align="center"|R0040
+
|align=center| 6
-
|align="center"|SpeI
+
-
|align="center"|PstI
+
-
|align="center"|2
+
|-
|-
-
|align="center"|D105
+
! rowspan="2" style="background: #D4E2EF;" | D104
-
|align="center"|MP105
+
! rowspan="2" style="background: #D4E2EF;" | J23109
-
|align="center"|S03154
+
|align=center|1
-
|align="center"|SpeI
+
! rowspan="2"| SpeI
-
|align="center"|PstI
+
! rowspan="2"| PstI
-
|align="center"|1
+
! rowspan="2"| BV
 +
! rowspan="2"| 2100 pb
 +
! rowspan="2"| 883 pb
 +
! rowspan="2"| 2000 pb
 +
! rowspan="2"| 700 pb
 +
! rowspan="1"| 1
 +
! rowspan="1"| 7
|-
|-
-
|align="center"|
+
|align=center|2
-
|align="center"|
+
! rowspan="1"| 1
-
|align="center"|
+
! rowspan="1"| 8
-
|align="center"|XbaI
+
-
|align="center"|PstI
+
-
|align="center"|2
+
|-
|-
-
|align="center"|
+
! rowspan="2" style="background: #D4E2EF;" | D105
-
|align="center"|
+
! rowspan="2" style="background: #D4E2EF;" | R0079
-
|align="center"|
+
|align=center|1
-
|align="center"|EcoRI
+
! rowspan="2"| SpeI
-
|align="center"|SpeI
+
! rowspan="2"| PstI
-
|align="center"|1
+
! rowspan="2"| BV
 +
! rowspan="2"| 2222 pb
 +
! rowspan="2"| 14 pb
 +
! rowspan="2"| 2000 pb
 +
! rowspan="2"| -
 +
|align=center| 1
 +
|align=center| 9
|-
|-
-
|align="center"|D106
+
|align=center|2
-
|align="center"|MP106
+
|align=center| 1
-
|align="center"|J23101
+
|align=center| 10
-
|align="center"|SpeI
+
-
|align="center"|PstI
+
-
|align="center"|1
+
|-
|-
-
|align="center"|
+
! rowspan="2" style="background: #D4E2EF;" | D106
-
|align="center"|
+
! rowspan="2" style="background: #D4E2EF;" | R0040
-
|align="center"|
+
|align=center|1
-
|align="center"|XbaI
+
! rowspan="2"| SpeI
-
|align="center"|PstI
+
! rowspan="2"| PstI
-
|align="center"|2
+
! rowspan="2"| BV
 +
! rowspan="2"| 2119 pb
 +
! rowspan="2"| 14 pb
 +
! rowspan="2"| 2000 pb
 +
! rowspan="2"| -
 +
|align=center| 1
 +
|align=center| 11
|-
|-
-
|align="center"|
+
|align=center|2
-
|align="center"|
+
|align=center| 1
-
|align="center"|
+
|align=center| 12
-
|align="center"|EcoRI
+
-
|align="center"|SpeI
+
-
|align="center"|1
+
|-
|-
-
|align="center"|D107
+
! rowspan="1" style="background: #D4E2EF;" | D107
-
|align="center"|MP107
+
! rowspan="1" style="background: #D4E2EF;" | S03154
-
|align="center"|C0079
+
|align=center|1
-
|align="center"|EcoRI
+
! rowspan="1"| SpeI
-
|align="center"|SpeI
+
! rowspan="1"| PstI
-
|align="center"|1
+
! rowspan="1"| BV
 +
! rowspan="1"| 2750 pb
 +
! rowspan="1"| 14 pb
 +
|align=center| 3000 pb
 +
|align=center| -
 +
|align=center| 2
 +
|align=center| 5
|-
|-
-
|align="center"|D108
+
! rowspan="2" style="background: #D4E2EF;" | D108
-
|align="center"|MP108
+
! rowspan="2" style="background: #D4E2EF;" | S03154
-
|align="center"|C0179
+
|align=center|2
-
|align="center"|EcoRI
+
! rowspan="2"| XbaI
-
|align="center"|SpeI
+
! rowspan="2"| PstI
-
|align="center"|1
+
! rowspan="2"| BI
 +
! rowspan="2"| 2057 pb
 +
! rowspan="2"| 707 pb
 +
! rowspan="2"| 2000 pb
 +
! rowspan="2"| 700 pb
 +
|align=center| 2
 +
|align=center| 2
|-
|-
-
|align="center"|D109
+
|align=center|3
-
|align="center"|MP109
+
|align=center| 2
-
|align="center"|E0030
+
|align=center| 3
-
|align="center"|EcoRI
+
-
|align="center"|SpeI
+
-
|align="center"|1
+
|-
|-
-
|align="center"|D110
+
! rowspan="1" style="background: #D4E2EF;" | D109
-
|align="center"|MP110
+
! rowspan="1" style="background: #D4E2EF;" | S03154
-
|align="center"|E0040
+
|align=center|4
-
|align="center"|EcoRI
+
| align=center|EcoRI
-
|align="center"|SpeI
+
| align=center|SpeI
-
|align="center"|1
+
| align=center|FI
 +
| align=center| 2056 pb
 +
| align=center| 708 pb
 +
|align=center| 2000 pb
 +
|align=center| 700 pb
 +
|align=center| 2
 +
|align=center| 4
|-
|-
-
|align="center"|D111
+
! rowspan="1" style="background: #D4E2EF;" | D110
-
|align="center"|MP111
+
! rowspan="1" style="background: #D4E2EF;" | S03879
-
|align="center"|E1010
+
|align=center|1
-
|align="center"|EcoRI
+
| align=center|SpeI
-
|align="center"|SpeI
+
| align=center|PstI
-
|align="center"|1
+
| align=center|BV
 +
| align=center| 2768 pb
 +
| align=center|14 pb
 +
|align=center| 2700 pb
 +
|align=center| -
 +
|align=center| 2
 +
|align=center| 6
|-
|-
-
|align="center"|D116
+
! rowspan="2" style="background: #D4E2EF;" | D111
-
|align="center"|MP116
+
! rowspan="2" style="background: #D4E2EF;" | S03879
-
|align="center"|J23100
+
|align=center|2
-
|align="center"|SpeI
+
! rowspan="2"| XbaI
-
|align="center"|PstI
+
! rowspan="2"| PstI
-
|align="center"|2
+
! rowspan="2"| BI
 +
! rowspan="2"| 2057 pb
 +
! rowspan="2"| 725 pb
 +
! rowspan="2"| 2000 pb
 +
! rowspan="2"| 700 pb
 +
|align=center| 2
 +
|align=center| 7
|-
|-
-
|align="center"|D117
+
|align=center|3
-
|align="center"|MP117
+
|align=center| 2
-
|align="center"|J23107
+
|align=center| 8
-
|align="center"|SpeI
+
-
|align="center"|PstI
+
-
|align="center"|2
+
|-
|-
-
|align="center"|D118
+
! rowspan="1" style="background: #D4E2EF;" | D112
-
|align="center"|MP118
+
! rowspan="1" style="background: #D4E2EF;" | S03879
-
|align="center"|B0015
+
|align=center|4
-
|align="center"|EcoRI
+
| align=center|EcoRI
-
|align="center"|XbaI
+
| align=center|SpeI
-
|align="center"|2
+
| align=center|FI
 +
| align=center| 2756 pb
 +
| align=center| 726 pb
 +
|align=center| 3000 pb
 +
|align=center| 700 pb
 +
|align=center| 2
 +
|align=center| 9
|-
|-
-
|align="center"|D119
+
! rowspan="1" style="background: #D4E2EF;" |D113
-
|align="center"|MP119
+
! rowspan="1" style="background: #D4E2EF;" |C0079
-
|align="center"|I0500
+
|align=center| 1
-
|align="center"|SpeI
+
|align=center|EcoRI
-
|align="center"|PstI
+
|align=center|SpeI
-
|align="center"|2
+
|align=center|FI
 +
|align=center| 4402 pb
 +
|align=center| 779 pb
 +
|align=center| 5000 pb
 +
|align=center| 800 pb
 +
|align=center| 2
 +
|align=center| 10
|-
|-
-
|align="center"|D120
+
! rowspan="1" style="background: #D4E2EF;" | D114
-
|align="center"|MP120
+
! rowspan="1" style="background: #D4E2EF;" | C0079
-
|align="center"|B0030
+
|align=center| 2
-
|align="center"|SpeI
+
|align=center|XbaI
-
|align="center"|PstI
+
|align=center|PstI
-
|align="center"|2
+
|align=center|BI
 +
|align=center| 4003 pb
 +
|align=center| 778 pb
 +
|align=center| 5000 pb
 +
|align=center| 800 pb
 +
|align=center| 2
 +
|align=center| 11
|-
|-
-
|align="center"|
+
! rowspan="1" style="background: #D4E2EF;"|D115
-
|align="center"|
+
! rowspan="1" style="background: #D4E2EF;"|C0179
-
|align="center"|
+
|align=center| 1
-
|align="center"|EcoRI
+
|align=center| EcoRI
-
|align="center"|XbaI
+
|align=center| SpeI
-
|align="center"|1
+
|align=center| FI
 +
|align=center| 4402 pb
 +
|align=center| 746 pb
 +
|align=center| 2000 pb
 +
|align=center| 800 pb
 +
|align=center| 2
 +
|align=center| 12
|-
|-
-
|align="center"|
+
! rowspan="1" style="background: #D4E2EF;" | D116
-
|align="center"|
+
! rowspan="1" style="background: #D4E2EF;" | C0179
-
|align="center"|
+
|align=center| 2
-
|align="center"|XbaI
+
|align=center| XbaI
-
|align="center"|PstI
+
|align=center| PstI
-
|align="center"|2
+
|align=center| BI
 +
|align=center| 4403 pb
 +
|align=center| 745 pb
 +
|align=center| 2000 pb
 +
|align=center| 800 pb
 +
|align=center| 3
 +
|align=center| 2
|-
|-
-
|align="center"|D121
+
! rowspan="1" style="background: #D4E2EF;"|D117
-
|align="center"|MP121
+
! rowspan="1" style="background: #D4E2EF;"|E0030
-
|align="center"|E0422
+
|align=center| 1
-
|align="center"|XbaI
+
|align=center|EcoRI
-
|align="center"|PstI
+
|align=center|SpeI
-
|align="center"|3
+
|align=center|FI
 +
|align=center| 3166 pb
 +
|align=center| 746 pb
 +
|align=center| 3000 pb
 +
|align=center| 700 pb
 +
|align=center| 3
 +
|align=center| 3
|-
|-
-
|align="center"|D122
+
! rowspan="1" style="background: #D4E2EF;" | D118
-
|align="center"|MP122
+
! rowspan="1" style="background: #D4E2EF;" | E0030
-
|align="center"|E0840
+
|align=center| 2
-
|align="center"|XbaI
+
|align=center| XbaI
-
|align="center"|PstI
+
|align=center| PstI
-
|align="center"|3
+
|align=center| BI
 +
|align=center| 3167 pb
 +
|align=center| 745 pb
 +
|align=center| 3000 pb
 +
|align=center| 700 pb
 +
|align=center| 3
 +
|align=center| 4
 +
|-
 +
! rowspan="1" style="background: #D4E2EF;"|D119
 +
! rowspan="1" style="background: #D4E2EF;"|E0040
 +
|align=center| 1
 +
|align=center| EcoRI
 +
|align=center| SpeI
 +
|align=center| FI
 +
|align=center| 2056 pb
 +
|align=center| 743 pb
 +
|align=center| 2000 pb
 +
|align=center| 700 pb
 +
|align=center| 3
 +
|align=center| 5
 +
|-
 +
! rowspan="1" style="background: #D4E2EF;" | D120
 +
! rowspan="1" style="background: #D4E2EF;" | E0040
 +
|align=center| 2
 +
|align=center| XbaI
 +
|align=center| PstI
 +
|align=center| BI
 +
|align=center| 2057 pb
 +
|align=center| 742 pb
 +
|align=center| 2000 pb
 +
|align=center| 700 pb
 +
|align=center| 3
 +
|align=center| 6
 +
|-
 +
! rowspan="1" style="background: #D4E2EF;"|D121
 +
! rowspan="1" style="background: #D4E2EF;"|E1010
 +
|align=center| 1
 +
|align=center| EcoRI
 +
|align=center| SpeI
 +
|align=center| FI
 +
|align=center| 4402 pb
 +
|align=center| 704 pb
 +
|align=center| 5000 pb
 +
|align=center| 700 pb
 +
|align=center| 3
 +
|align=center| 7
 +
|-
 +
! rowspan="1" style="background: #D4E2EF;" | D122
 +
! rowspan="1" style="background: #D4E2EF;" | E1010
 +
|align=center| 2
 +
|align=center| XbaI
 +
|align=center| PstI
 +
|align=center| BI
 +
|align=center| 4403 pb
 +
|align=center| 703 pb
 +
|align=center| 5000 pb
 +
|align=center| 700 pb
 +
|align=center| 3
 +
|align=center| 8
 +
|-
 +
! rowspan="2" style="background: #D4E2EF;"|D123
 +
! rowspan="2" style="background: #D4E2EF;"|J23100
 +
|align=center|1
 +
! rowspan="2"| SpeI
 +
! rowspan="2"| PstI
 +
! rowspan="2"| BV
 +
! rowspan="2"| 2100 pb
 +
! rowspan="2"| 883 pb
 +
! rowspan="2"| 2000 pb
 +
! rowspan="2"| 900 pb
 +
! rowspan="1"| 3
 +
! rowspan="1"| 9
 +
|-
 +
|align=center|2
 +
! rowspan="1"| 3
 +
! rowspan="1"| 10
 +
|-
 +
! rowspan="2" style="background: #D4E2EF;"|D124
 +
! rowspan="2" style="background: #D4E2EF;"|J23107
 +
|align=center|1
 +
! rowspan="2"| SpeI
 +
! rowspan="2"| PstI
 +
! rowspan="2"| BV
 +
! rowspan="2"| 2100 pb
 +
! rowspan="2"| 883 pb
 +
! rowspan="2"| 2000 pb
 +
! rowspan="2"| 900 pb
 +
! rowspan="1"| 3
 +
! rowspan="1"| 11
 +
|-
 +
|align=center|2
 +
! rowspan="1"| 3
 +
! rowspan="1"| 12
 +
|-
 +
! rowspan="2" style="background: #D4E2EF;"|D125
 +
! rowspan="2" style="background: #D4E2EF;"|B0015
 +
|align=center|1
 +
! rowspan="2"|EcoRI
 +
! rowspan="2"|XbaI
 +
! rowspan="2"| BV
 +
! rowspan="2"| 3303 pb
 +
! rowspan="2"| 15 pb
 +
! rowspan="2"| 3000 pb
 +
! rowspan="2"| -
 +
! rowspan="1"| 4
 +
! rowspan="1"| 2
 +
|-
 +
|align=center|2
 +
! rowspan="1"| 4
 +
! rowspan="1"| 3
 +
|-
 +
! rowspan="2" style="background: #D4E2EF;"|D126
 +
! rowspan="2" style="background: #D4E2EF;"|I0500
 +
|align=center|1
 +
! rowspan="2"|SpeI
 +
! rowspan="2"|PstI
 +
! rowspan="2"| FV
 +
! rowspan="2"| 5621 pb
 +
! rowspan="2"| 14 pb 
 +
! rowspan="2"| 6000 pb
 +
! rowspan="2"| -
 +
! rowspan="1"| 4
 +
! rowspan="1"| 4
 +
|-
 +
|align=center|2
 +
! rowspan="1"| 4
 +
! rowspan="1"| 5
 +
|-
 +
! rowspan="2" style="background: #D4E2EF;" | D127
 +
! rowspan="2" style="background: #D4E2EF;" | B0030
 +
|align=center|1
 +
! rowspan="2"| XbaI
 +
! rowspan="2"| PstI
 +
! rowspan="2"| BI
 +
! rowspan="2"| 2057 pb
 +
! rowspan="2"| 37 pb
 +
! rowspan="2"| 2000 pb
 +
! rowspan="2"| 50 pb
 +
! rowspan="1"| 6
 +
! rowspan="1"| 4
 +
|-
 +
|align=center|2
 +
! rowspan="1"| 6
 +
! rowspan="1"| 5
 +
|-
 +
! rowspan="1" style="background: #D4E2EF;" | D128
 +
! rowspan="1" style="background: #D4E2EF;" | B0030
 +
|align=center|3
 +
! rowspan="1"|  EcoRI
 +
! rowspan="1"|  XbaI
 +
! rowspan="1"|  FV
 +
! rowspan="1"| 2079 pb
 +
! rowspan="1"|  15 pb
 +
! rowspan="1"| 2200 pb
 +
! rowspan="1"| -
 +
! rowspan="1"| 4
 +
! rowspan="1"| 6
 +
|-
 +
! rowspan="2" style="background: #D4E2EF;" | D129
 +
! rowspan="2" style="background: #D4E2EF;" | B0030
 +
|align=center|4
 +
! rowspan="2"| SpeI
 +
! rowspan="2"| PstI
 +
! rowspan="2"| BV
 +
! rowspan="2"| 2080 pb
 +
! rowspan="2"| 14 pb
 +
! rowspan="2"| 2000 pb
 +
! rowspan="2"| -
 +
! rowspan="1"| 4
 +
! rowspan="1"| 7
 +
|-
 +
|align=center|5
 +
! rowspan="1"| 4
 +
! rowspan="1"| 8
 +
|-
 +
! rowspan="3" style="background: #D4E2EF;"|D130
 +
! rowspan="3" style="background: #D4E2EF;"|E0422
 +
|align=center|1
 +
! rowspan="3"| XbaI
 +
! rowspan="3"| PstI
 +
! rowspan="3"| BI
 +
! rowspan="3"| 2057 pb
 +
! rowspan="3"| 939 pb
 +
! rowspan="3"| 2000 pb
 +
! rowspan="3"| 1100 pb
 +
! rowspan="1"|  4
 +
! rowspan="1"|  9
 +
|-
 +
|align=center|2
 +
! rowspan="1"|  4
 +
! rowspan="1"|  10
 +
|-
 +
|align=center|3
 +
! rowspan="1"| 4
 +
! rowspan="1"| 11
 +
|-
 +
! rowspan="3" style="background: #D4E2EF;"|D131
 +
! rowspan="3" style="background: #D4E2EF;"|E0840
 +
|align=center|1
 +
! rowspan="3"| XbaI
 +
! rowspan="3"| PstI
 +
! rowspan="3"| BI
 +
! rowspan="3"| 2057 pb
 +
! rowspan="3"| 900 pb
 +
! rowspan="1"| 2000 pb
 +
! rowspan="1"| 1000 pb
 +
! rowspan="1"|  4
 +
! rowspan="1"|  12
 +
|-
 +
|align=center|2
 +
! rowspan="2"| 2000 pb
 +
! rowspan="2"| 900 pb
 +
! rowspan="1"|  5
 +
! rowspan="1"|  2
 +
|-
 +
|align=center|3
 +
! rowspan="1"| 5
 +
! rowspan="1"| 3
|}
|}
-
== Results of digestions : Electrophoresis ==
+
 
 +
==> conclusion : all the digestion have succeed.....GREAT !
 +
 
 +
== DNA extraction  ==
 +
 
 +
 
 +
* Cutting of the parts of interest, for all the digestion that have migrated on the gels
 +
* Use of Promega's protocol for the extraction.
 +
* Test of the succeed of the extraction by electrophoresis on 2µl of the parts extracted.
 +
 
 +
 
 +
==> conclusion : we succeed to detect DNA in our samples this time.

Latest revision as of 16:55, 31 July 2008

← Yesterday

↓ Calendar ↑

Tomorrow →


Contents

MiniPreps

  • Use of Promega's protocol on all the clones cultivated on the 25th.
  • Preparation of 50µl of minipreps in double.


Name Biobrick Description
MP100 [http://partsregistry.org/Part:BBa_B0034 B0034] Strongest RBS (Efficiency = 1)
MP101 [http://partsregistry.org/Part:BBa_J23101 J23101] Strongest Constitutive promoter
MP102 [http://partsregistry.org/Part:BBa_J23109 J23109] Weak Constitutive promoter
MP103 [http://partsregistry.org/Part:BBa_R0079 R0079] promoter pLas (lasr & PAI regulated)
MP104 [http://partsregistry.org/Part:BBa_R0040 R0040] tetR repressible promoter
MP105 [http://partsregistry.org/Part:BBa_S03154 S03154] B0034(rbs) ->LasI
MP106 [http://partsregistry.org/Part:BBa_S03879 S03879] B0034(rbs) -> TetR
MP107 [http://partsregistry.org/Part:BBa_C0079 C0079] lasR activator with LVA Tag
MP108 [http://partsregistry.org/Part:BBa_C0179 C0179] lasR activator
MP109 [http://partsregistry.org/Part:BBa_E0030 E0030] YFP
MP110 [http://partsregistry.org/Part:BBa_E0040 E0040] GFP
MP111 [http://partsregistry.org/Part:BBa_E1010 E1010] mRFP
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°6 : 100 pb)
  • 30µl of digestion added with 5µl of loading Dye 6x
  • migration ~30min at 100W
  • Gel 1, 2, 3, 4, 5 = 0.8%
  • Gel 6 = 1,5%


gel1 gel1 gel2 gel2 gel3 gel3 gel4 gel4


gel5 gel5 gel6 gel6


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 1800 pb 40 pb 6 2
2 6 3
D101 B0034 3 EcoRI XbaI FV 2076 pb 15 pb 2000 pb - 1 2
D102 B0034 4 SpeI PstI BV 2077 pb 14 pb 2000 pb - 1 3
5 1 4
D103 J23101 1 SpeI PstI BV 2100 pb 883 pb 2000 pb 700 pb 1 5
2 1 6
D104 J23109 1 SpeI PstI BV 2100 pb 883 pb 2000 pb 700 pb 1 7
2 1 8
D105 R0079 1 SpeI PstI BV 2222 pb 14 pb 2000 pb - 1 9
2 1 10
D106 R0040 1 SpeI PstI BV 2119 pb 14 pb 2000 pb - 1 11
2 1 12
D107 S03154 1 SpeI PstI BV 2750 pb 14 pb 3000 pb - 2 5
D108 S03154 2 XbaI PstI BI 2057 pb 707 pb 2000 pb 700 pb 2 2
3 2 3
D109 S03154 4 EcoRI SpeI FI 2056 pb 708 pb 2000 pb 700 pb 2 4
D110 S03879 1 SpeI PstI BV 2768 pb 14 pb 2700 pb - 2 6
D111 S03879 2 XbaI PstI BI 2057 pb 725 pb 2000 pb 700 pb 2 7
3 2 8
D112 S03879 4 EcoRI SpeI FI 2756 pb 726 pb 3000 pb 700 pb 2 9
D113 C0079 1 EcoRI SpeI FI 4402 pb 779 pb 5000 pb 800 pb 2 10
D114 C0079 2 XbaI PstI BI 4003 pb 778 pb 5000 pb 800 pb 2 11
D115 C0179 1 EcoRI SpeI FI 4402 pb 746 pb 2000 pb 800 pb 2 12
D116 C0179 2 XbaI PstI BI 4403 pb 745 pb 2000 pb 800 pb 3 2
D117 E0030 1 EcoRI SpeI FI 3166 pb 746 pb 3000 pb 700 pb 3 3
D118 E0030 2 XbaI PstI BI 3167 pb 745 pb 3000 pb 700 pb 3 4
D119 E0040 1 EcoRI SpeI FI 2056 pb 743 pb 2000 pb 700 pb 3 5
D120 E0040 2 XbaI PstI BI 2057 pb 742 pb 2000 pb 700 pb 3 6
D121 E1010 1 EcoRI SpeI FI 4402 pb 704 pb 5000 pb 700 pb 3 7
D122 E1010 2 XbaI PstI BI 4403 pb 703 pb 5000 pb 700 pb 3 8
D123 J23100 1 SpeI PstI BV 2100 pb 883 pb 2000 pb 900 pb 3 9
2 3 10
D124 J23107 1 SpeI PstI BV 2100 pb 883 pb 2000 pb 900 pb 3 11
2 3 12
D125 B0015 1 EcoRI XbaI BV 3303 pb 15 pb 3000 pb - 4 2
2 4 3
D126 I0500 1 SpeI PstI FV 5621 pb 14 pb 6000 pb - 4 4
2 4 5
D127 B0030 1 XbaI PstI BI 2057 pb 37 pb 2000 pb 50 pb 6 4
2 6 5
D128 B0030 3 EcoRI XbaI FV 2079 pb 15 pb 2200 pb - 4 6
D129 B0030 4 SpeI PstI BV 2080 pb 14 pb 2000 pb - 4 7
5 4 8
D130 E0422 1 XbaI PstI BI 2057 pb 939 pb 2000 pb 1100 pb 4 9
2 4 10
3 4 11
D131 E0840 1 XbaI PstI BI 2057 pb 900 pb 2000 pb 1000 pb 4 12
2 2000 pb 900 pb 5 2
3 5 3


==> conclusion : all the digestion have succeed.....GREAT !

DNA extraction

  • Cutting of the parts of interest, for all the digestion that have migrated on the gels
  • Use of Promega's protocol for the extraction.
  • Test of the succeed of the extraction by electrophoresis on 2µl of the parts extracted.


==> conclusion : we succeed to detect DNA in our samples this time.

Retrieved from "http://2008.igem.org/Team:Paris/July_26"