Difference between revisions of "Team:Paris Saclay/Notebook/August/12"
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Agarose gel 1%, Migration 90V | Agarose gel 1%, Migration 90V | ||
− | + | [[File:ParisSaclay 12.08.15-quantif.jpg|300px|center]] | |
+ | <html><p><i>Quantification, from left to right: 1. <a href="https://2015.igem.org/File:Paris_Saclay-Ladder.jpg" target="_blank">DNA Ladder</a>, 2. BBa_K1707020, 3. BBa_K1707030, 4. BBa_I13602#1, 5. BBa_I13602#2, 6. Empty, 7. Empty, 8. Empty, 9. Empty, 10. Empty</i></p></html> | ||
We can conclude: | We can conclude: | ||
* BBa_I13602 #1: 40 µg/µL | * BBa_I13602 #1: 40 µg/µL | ||
Line 43: | Line 44: | ||
* BBa_K1707011 #3 | * BBa_K1707011 #3 | ||
* BBa_K1707012 #2 | * BBa_K1707012 #2 | ||
+ | [[File:ParisSaclay 12.08.15-vérif purif.jpg|300px|center]] | ||
+ | <html><p><i>Verification of gel purification, from left to right: 1. <a href="https://2015.igem.org/File:Paris_Saclay-Ladder.jpg" target="_blank">DNA Ladder</a>, 2. BBa_K1707011#3, 3. BBa_K1707012#2, 4. Empty, 5. Empty, 6. Empty, 7. Empty, 8. Empty, 9. Empty, 10. Empty</i></p></html> | ||
We can conclude that biobricks are OK | We can conclude that biobricks are OK |
Latest revision as of 20:15, 18 September 2015
Contents
Wednesday 12th August
Lab Work
Purification
by Pauline
- BBa_I13602 #1
- BBa_I13602 #2
- BBa_K1707020
- BBa_K1707030
With Macherey-Nagel Purification kit
Quantification
by Pauline
Agarose gel 1%, Migration 90V
Quantification, from left to right: 1. DNA Ladder, 2. BBa_K1707020, 3. BBa_K1707030, 4. BBa_I13602#1, 5. BBa_I13602#2, 6. Empty, 7. Empty, 8. Empty, 9. Empty, 10. Empty
We can conclude:- BBa_I13602 #1: 40 µg/µL
- BBa_I13602 #2: 40 µg/µL
- BBa_K1707020: 10 µg/µL
- BBa_K1707030: 13 µg/µL
Digestion
by Coralie
- BBa_K1707011 #3 and BBa_K1707012 #2
- Mix for each reaction:
- 1 µL PstI
- 1 µL XbaI
- 2 µL Buffer FastDigest 10x
- 6 µL H2O
- 10 µL plasmid
- Incubation 37°C, 2h
- Mix for each reaction:
Purification on gel
by Pauline
Agarose gel 1%, 90V
- BBa_K1707011 #3
- BBa_K1707012 #2
Verification of gel purification, from left to right: 1. DNA Ladder, 2. BBa_K1707011#3, 3. BBa_K1707012#2, 4. Empty, 5. Empty, 6. Empty, 7. Empty, 8. Empty, 9. Empty, 10. Empty
We can conclude that biobricks are OK
Purification by the Macherey-Nagel Purification kit
Ligation
by Pauline
- BBa_K1707035: BBa_K1707030 + BBa_I13602
- 11,6 µL BBa_K1707030
- 1,25 µL BBa_R0040
- 2 µL Ligase
- 2 µL Buffer 10x
- 3,15 µL H2O
- BBa_K1707036: BBa_K1707020 + BBa_I13602
- 15,5 µL BBa_K1707006
- 1,25 µL BBa_B0030
- 2 µL Ligase
- 2,5 µL Buffer 10x
- 3,75 µL H2O
Incubation 4°C, ON
New culture
by Pauline
- From stock:
- BBa_K1707012 #2 x2
- BBa_K1707011 #3 x2
- BBa_R0051
- Two clones from plate of each:
- BBa_K1707033
- BBa_K1707031
- BBa_K1707016
In 5mL LB + 5µL of appropriate antibiotic
Soil experiment
Member present:
- Instructors: Claire
- Students: Coralie and Pauline