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MiniPrep
The protocol of MiniPrep of Quiagen has been done on all the clones cultivated on the 18th.
J23109 clones 1,2,5
E0030 clones 1,2,3
pSB1A3 clones 1,2,3
C0179 clones 1,2,7
C0079 clones 1,2,3
R0079 clones 3,4,6
E1010 clones 1,2,3
B0034 clones 1,2,3
J23101 clones 1,2,3
E0040 clones 1,2
R0040 clones 1,2
We did an error during the manipulation (probably an exchange between some tubes) so we decided to screen again clones of B0034 and R0040.
Quantitative analysis of ADN
gel 1
| Band
| Biobricks
| Clone
| Expected size
| Measured size
|
| 1
| Ladder 1 kpb
|
|
|
|
| 2
| E1010
| 1
| 5.106 kb
| ~6-8 kb
|
| 3
| E1010
| 2
| 5.106 kb
| ~6-8 kb
|
| 4
| C0079
| 1
| 5.181 kb
| ~6-8 kb
|
| 5
| C0079
| 2
| 5.181 kb
| ~6-8 kb
|
| 6
| C0079
| 3
| 5.181 kb
| ~6-8 kb
|
| 7
| J23101 : problem !!!
| 1
| 3.037 kb
| ~6-8 kb
|
| 8
| J23101 : problem !!!
| 2
| 3.037 kb
| ~6-8 kb
|
| 9
| J23101 : problem !!!
| 3
| 3.037 kb
| ~6-8 kb
|
| 10
| B0034
| 2
| 2.091 kb
| ~2 kb
|
| 11
| B0034
| X
| 2.091 kb
| ~2 kb
|
| 12
| B0034 : problem !!!
| Y
| 2.091 kb
| ~5 kb
|
| 13
| E0040
| 1
| 2.799 kb
| ~2.5-3 kb
|
| 14
| E0040
| 2
| 2.799 kb
| ~2.5-3 kb
|
| 15
| R0040
| ?
| 2.133 kb
| ~2 kb
|
We see bad results for B0034 (bands 10 to 12), R0040 (band 15). We'll try again!
gel 2
| Band
| Biobricks
| Clone
| Expected size
| Measured size
|
| 1
| Ladder 1 kpb
|
|
|
|
| 2
| J23109
| 1
| 3.037 kb
| ~ 2-3 kb
|
| 3
| J23109 : Problem !!!
| 2
| 3.037 kb
| ~ 6-8 kb
|
| 4
| J23109 : Problem !!!
| 3
| 3.037 kb
| > 10 kb
|
| 5
| pSB1A3
| 1
| 2.157 kb
| ~ 2-3 kb
|
| 6
| pSB1A3
| 2
| 2.157 kb
| ~ 2-3 kb
|
| 7
| pSB1A3
| 3
| 2.157 kb
| ~ 2-3 kb
|
| 8
| E0030
| 1
| 3.912 kb
| ~ 3.5 kb
|
| 9
| E0030
| 2
| 3.912 kb
| ~ 3.5 kb
|
| 10
| E0030
| 3
| 3.912 kb
| ~ 3.5 kb
|
| 11
| R0079
| 3
| 2.236 kb
| ~ 1.5 kb
|
| 12
| R0079
| 4
| 2.236 kb
| ~ 1.5 kb
|
| 13
| R0079
| 6
| 2.236 kb
| ~ 1.5 kb
|
| 14
| C0179 : Problem !!!
| 1
| 2.802 kb
| > 10 kb
|
| 15
| C0179 : Problem !!!
| 2
| 2.802 kb
| ~ 10 kb
|
| 16
| C0179
| 3
| 2.802 kb
| ~ 2.5 kb
|
We see bad results for J23109 (bands 1 to 3), C0179 (bands 14 to 16). We'll try again!
Screening R0040 and B0034
PCR
For 1 tube :
25µL Quick Load
1µL Oligo F
1µL Oligo R
23µL H2O
Program SCREENIN
Electrophoresis
| Band
| Biobricks
| Clone
| Expected size
| Measured size
|
| 2
| Ladder 100 pb
|
|
|
|
| 3
| B0034
| 1
| 250 pb
| nothing
|
| 4
| B0034
| 2
| 250 pb
| ~250pb
|
| 5
| R0040
| 1
| 292 pb
| ~300pb
|
| 6
| R0040 : Problem !!!
| 2
| 292 pb
| ~250pb
|
| 7
| R0040 : Problem !!!
| ?
| 292 pb
| ~250pb
|
|
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