Difference between revisions of "Team:Paris Saclay/Notebook/July/9"

(Electrophoresis)
(Electrophoresis)
 
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* BBa_J23106 + BBa_I13504
 
* BBa_J23106 + BBa_I13504
 
* BBa_J23117 + BBa_I13504
 
* BBa_J23117 + BBa_I13504
On LB + Ampicillin 100ug/mL.
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On LB + Chloramphenicol 20ug/mL.
 
Incubation ON, 37°C
 
Incubation ON, 37°C
  
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Migration 0,06A 80V
 
Migration 0,06A 80V
  
[[File:ParisSaclay_09072015_-_Quantification.jpg|thumb|]]
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[[File:ParisSaclay_09072015_-_Quantification.jpg|300px|center]]
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<html><i><p>Quantification, from left to right: 1. <a href="https://2015.igem.org/File:Paris_Saclay-Ladder.jpg" target="_blank">DNA Ladder</a>, 2. BBa_C0040, 3. BBa_K115017, 4. BBa_J23101, 5. Empty, 6. Empty, 7. Empty, 8. Empty, 9. Empty, 10. Empty, 11. Empty, 12. Empty</i></p></i></html>
  
 
===Quantification===
 
===Quantification===

Latest revision as of 21:14, 18 September 2015


Thursday 9th July

Lab Work

Transformation

by Coralie

Ligation product:

  • BBa_J23101 + BBa_I13504
  • BBa_J23106 + BBa_I13504
  • BBa_J23117 + BBa_I13504

On LB + Chloramphenicol 20ug/mL. Incubation ON, 37°C

by Johan and Seong Ko

  • BBa_R0051

On LB + Ampicillin 100ug/mL. Incubation ON, 37°C

Plasmid Rehydratation

by Johan and Audrey

  • BBa_S03518
  • BBa_B0030
  • BBa_B0015
  • BBa_K1399005

Digestion

by Pauline and Audrey

Plasmid with promotor: BBa_J23101

  • 10µL of our plasmid with promotor
  • 1µL SpeI
  • 1µL PstI
  • 2µL buffer 10x FD

Plasmid with gene: BBa_COO40 and BBa_K115017

  • 10µL of our plasmid with gene
  • 1µL XbaI
  • 1µL PstI
  • 2µL buffer 10x FD

After this step, we separate the sequence we need from the sequence we don't with electrophoresis

Electrophoresis

by Pauline and Audrey

Preparation of Agarose Gel 1%, 0,5g in 50mL of 1X TAE, 0,5µL of BET Migration 0,06A 80V

ParisSaclay 09072015 - Quantification.jpg

Quantification, from left to right: 1. DNA Ladder, 2. BBa_C0040, 3. BBa_K115017, 4. BBa_J23101, 5. Empty, 6. Empty, 7. Empty, 8. Empty, 9. Empty, 10. Empty, 11. Empty, 12. Empty

Quantification

by Pauline and Audrey

Preparation of Agarose Gel 1%, 0,5g in 50mL of 1X TAE, 0,5µL of BET Migration 0,06A 80V We quantify with the electrophoresis gel:

  • BBa_C0040: 10ng/µL
  • BBa_J23101: 20ng/µL
  • BBa_K115017: we don't see enough of fluorescent to quantify the biobrick. We decide to amplify this one by PCR.

Members present:

  • Instructors and advisors: Alice.
  • Students: Johan, Seong Koo, Audrey, Coralie, Pauline

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