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conditions :
- 10µl of ladder 1 kb
- xµl of extraction added with 2µl of loading Dye 6x
- migration ~30min at 100W
- Gel 1, 2, 3, 4 = 0,8%
- 2µl of extraction added with 2µl of loading Dye 6x
- Gel 5, 6 = 0,8%
- 3µl of extraction added with 2µl of loading Dye 6x
- Gel 7 = 0,8%
- 5µl of extraction added with 2µl of loading Dye 6x
- Gel 8 = 1,5%
- 10µl of extraction added with 2µl of loading Dye 6x
gel1
gel2
gel3
gel4
gel5
gel6
gel7
gel8
Name
| BioBrick
| Tube N°
| Enz 1
| Enz 2
| Obs
| Exp Size
| Mea Size
| Conc (ng/µl)
| Gel
| Band
|
D100
| B0034
| 1
| XbaI
| PstI
| BI
| 34 pb
| -
| -
| 8
| 5
|
2
| -
| -
| -
|
D101
| B0034
| 3
| EcoRI
| XbaI
| FV
| 2076 pb
| - & 2000 pb
| - & 7
| 1 & 7
| 2 & 2
|
D102
| B0034
| 4
| SpeI
| PstI
| BV
| 2077 pb
| 2000 pb
| - & 5
| 1 & 7
| 3 & 3
|
5
| - & 10
| 1 & 7
| 4 & 4
|
D103
| J23101
| 1
| SpeI
| PstI
| BV
| 2100 pb
| 2000 pb
| -
| 1
| 5
|
2
| 10
| 1
| 6
|
D104
| J23109
| 1
| SpeI
| PstI
| BV
| 2100 pb
| 2000 pb
| 20
| 1
| 7
|
2
| 20
| 1
| 8
|
D105
| R0079
| 1
| SpeI
| PstI
| BV
| 2222 pb
| 2000 pb
| 5
| 2
| 2
|
2
| 5
| 2
| 3
|
D106
| R0040
| 1
| SpeI
| PstI
| BV
| 2119 pb
| - & 2000 pb
| - & 5
| 2 & 7
| 4 & 5
|
2
| - & 8
| 2 & 7
| 5 & 6
|
D107
| S03154
| 1
| SpeI
| PstI
| BV
| 2750 pb
| - & 3000 pb
| - & 5
| 2 & 7
| 6 & 7
|
D108
| S03154
| 2
| XbaI
| PstI
| BI
| 707 pb
| - & -
| - & -
| 2 & 7
| 7 & 8
|
3
| - & -
| 2 & 7
| 8 & 9
|
D109
| S03154
| 4
| EcoRI
| SpeI
| FI
| 708 pb
| - & -
| - & -
| 3 & 7
| 2 & 10
|
D110
| S03879
| 1
| SpeI
| PstI
| BV
| 2768 pb
| 3000 pb
| 20
| 3
| 3
|
D111
| S03879
| 2
| XbaI
| PstI
| BI
| 725 pb
| -
| -
| 3
| 4
|
3
| -
| 3
| 5
|
D112
| S03879
| 4
| EcoRI
| SpeI
| FI
| 726 pb
| -
| -
| 3
| 6
|
D113
| C0079
| 1
| EcoRI
| SpeI
| FI
| 779 pb
| 800 pb
| 20
| 3
| 7
|
D114
| C0079
| 2
| XbaI
| PstI
| BI
| 778 pb
| 800 pb
| 20
| 3
| 8
|
D115
| C0179
| 1
| EcoRI
| SpeI
| FI
| 746 pb
| - & -
| - & -
| 4 & 7
| 2 & 11
|
D116
| C0179
| 2
| XbaI
| PstI
| BI
| 745 pb
| - & -
| - & -
| 4 & 7
| 3 & 12
|
D117
| E0030
| 1
| EcoRI
| SpeI
| FI
| 746 pb
| 700 pb
| 5
| 4
| 4
|
D118
| E0030
| 2
| XbaI
| PstI
| BI
| 745 pb
| 700 pb
| 5
| 4
| 5
|
D119
| E0040
| 1
| EcoRI
| SpeI
| FI
| 743 pb
| - & 800 pb
| - & 10
| 4 & 7
| 6 & 13
|
D120
| E0040
| 2
| XbaI
| PstI
| BI
| 742 pb
| - & 800 pb
| - & 10
| 4 & 7
| 7 & 14
|
D121
| E1010
| 1
| EcoRI
| SpeI
| FI
| 704 pb
| 700 pb
| 5
| 4
| 8
|
D122
| E1010
| 2
| XbaI
| PstI
| BI
| 703 pb
| 800 pb
| 10
| 5
| 2
|
D123
| J23100
| 1
| SpeI
| PstI
| BV
| 2100 pb
| 2000 pb
| 10
| 5
| 3
|
2
| 10
| 5
| 4
|
D124
| J23107
| 1
| SpeI
| PstI
| BV
| 2100 pb
| 2000 pb
| 10
| 5
| 5
|
2
| 15
| 5
| 6
|
D124
| J23107
| 1
| EcoRI
| XbaI
| FV
| 3303 pb
| 3000 pb
| 15
| 5
| 7
|
2
| 15
| 5
| 8
|
D126
| I0500
| 1
| SpeI
| PstI
| FV
| 5621 pb
| 6000 pb
| 10
| 5
| 9
|
2
| 10
| 5
| 10
|
D127
| B0030
| 1
| XbaI
| PstI
| BI
| 37 pb
| - & -
| - & -
| 8
| 3
|
2
| -
| -
| -
|
D128
| B0030
| 3
| EcoRI
| XbaI
| FV
| 2079 pb
| 2000 pb
| 10
| 5
| 11
|
D129
| B0030
| 4
| SpeI
| PstI
| BV
| 2080 pb
| 2000 pb
| 10
| 5
| 12
|
5
| 10
| 5
| 13
|
D130
| E0422
| 1
| XbaI
| PstI
| BI
| 939 pb
| 1100 pb
| 10
| 5
| 14
|
2
| 10
| 5
| 15
|
3
| 10
| 5
| 16
|
D131
| E0840
| 1
| XbaI
| PstI
| BI
| 900 pb
| 1000 pb
| 5
| 6
| 3
|
2
| 10
| 6
| 4
|
3
| 5
| 6
| 5
|
==> Conclusion : for most of the samples we have enough signal to determine the concentration of the digestion to do the ligation.
For the samples we don't succeed to obtain signals, we have migrated a second gel ( gel n°7).
==> Conclusion : The gel n°7 allowed us to know the concentration of other digestion undetermined.
Ligation
Conditions :
Name
| Vector
| Insert
| Antibio
| Vol. vector (µl)
| Vol. insert (µl)
| Vol. H20 (µl)
|
L100
| D110
| D130
| A
| 3
| 6
| 8
|
L101
| D110
| D131
| A
| 3
| 6
| 8
|
L102
| D129
| D1118
| A
| 5
| 10
| 2
|
L103
| D129
| D122
| A
|
|
|
|
L104
| D129
| D114
| A
|
|
|
|
L105
| D123
| D130
| A
|
|
|
|
L106
| D123
| D131
| A
|
|
|
|
L107
| D103
| D130
| A
|
|
|
|
L108
| D103
| D131
| A
|
|
|
|
L109
| D124
| D130
| A
|
|
|
|
L110
| D124
| D131
| A
|
|
|
|
|