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

(Thursday 9th July)
(Electrophoresis)
 
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{{Team:Paris_Saclay/notebook_header}}
 
=Thursday 9th July=
 
=Thursday 9th July=
 
==Lab Work==
 
==Lab Work==
====Transformation====
+
===Transformation===
 
''by Coralie''
 
''by Coralie''
  
 
Ligation product:
 
Ligation product:
* J23101 + I13504
+
* BBa_J23101 + BBa_I13504
* J23106 + I13504
+
* BBa_J23106 + BBa_I13504
* J23117 + I13504
+
* BBa_J23117 + BBa_I13504
On LB + Ampicillin 100ug/mL  
+
On LB + Chloramphenicol 20ug/mL.
 
Incubation ON, 37°C
 
Incubation ON, 37°C
 
  
 
''by Johan and Seong Ko''
 
''by Johan and Seong Ko''
 
+
* BBa_R0051
R0051
+
On LB + Ampicillin 100ug/mL.
On LB + Ampicillin 100ug/mL  
+
 
Incubation ON, 37°C
 
Incubation ON, 37°C
  
===Plasmid Rehydratation :===
+
===Plasmid Rehydratation===
 
''by Johan and Audrey''
 
''by Johan and Audrey''
  
Biobricks:
+
* BBa_S03518
* S03518
+
* BBa_B0030
* B0030
+
* BBa_B0015
* B0015
+
* BBa_K1399005
* K1399005
+
  
====Digestion====
+
===Digestion===
 
''by Pauline and Audrey''
 
''by Pauline and Audrey''
 
   
 
   
Plasmid with promotor: J23101
+
Plasmid with promotor: BBa_J23101
 
* 10µL of our plasmid with promotor
 
* 10µL of our plasmid with promotor
 
* 1µL SpeI
 
* 1µL SpeI
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* 2µL buffer 10x FD
 
* 2µL buffer 10x FD
  
Plasmid with gene: COO40 and K115017
+
Plasmid with gene: BBa_COO40 and BBa_K115017
 
* 10µL of our plasmid with gene
 
* 10µL of our plasmid with gene
 
* 1µL XbaI
 
* 1µL XbaI
Line 44: Line 42:
 
After this step, we separate the sequence we need from the sequence we don't with electrophoresis
 
After this step, we separate the sequence we need from the sequence we don't with electrophoresis
  
====Electrophoresis====
+
===Electrophoresis===
 
''by Pauline and Audrey''
 
''by Pauline and Audrey''
  
 
Preparation of Agarose Gel 1%, 0,5g in 50mL of 1X TAE, 0,5µL of BET
 
Preparation of Agarose Gel 1%, 0,5g in 50mL of 1X TAE, 0,5µL of BET
Migration 0,06A 80V  
+
Migration 0,06A 80V
  
====Quantification====
+
[[File:ParisSaclay_09072015_-_Quantification.jpg|300px|center]]
 +
<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===
 
''by Pauline and Audrey''
 
''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:
 
We quantify with the electrophoresis gel:
* C0040: 10ng/µL
+
* BBa_C0040: 10ng/µL
* J23101: 20ng/µL
+
* BBa_J23101: 20ng/µL
* K115017: we don't see enough of fluorescent to quantify the biobrick. We decide to amplify this one by PCR.
+
* BBa_K115017: we don't see enough of fluorescent to quantify the biobrick. We decide to amplify this one by PCR.
  
 
'''Members present:'''
 
'''Members present:'''
 
* Instructors and advisors: Alice.
 
* Instructors and advisors: Alice.
 
* Students: Johan, Seong Koo, Audrey, Coralie, Pauline
 
* Students: Johan, Seong Koo, Audrey, Coralie, Pauline
 +
{{Team:Paris_Saclay/notebook_footer}}

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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