Difference between revisions of "Team:Paris Saclay/Notebook/July/8"

(→‎Wednesday 8th July)
(→‎Wednesday 8th July)
Line 4: Line 4:
 
''by Audrey''
 
''by Audrey''
  
−
R0051  
+
Biobrick: R0051 (2015)
 
On LB + Ampicillin 100ug/mL  
 
On LB + Ampicillin 100ug/mL  
  
Line 10: Line 10:
 
''by Seong Koo, Johan''
 
''by Seong Koo, Johan''
  
−
R0051 from different years
+
R0051 from different years (2013 and 2014)
  
 
====Digestion====
 
====Digestion====
 
''by Pauline, Coralie''
 
''by Pauline, Coralie''
  
−
10uL of our plasmid with promotor
+
First Digestion:
−
1uL SpeI
+
* Biobricks:
−
1uL PstI
+
** J23101
−
2uL buffer 10x FD
+
** J23106
 +
** J23117
  
−
*J23101
+
* Mix:
−
*J23106
+
** 10µL of our plasmid with promotor
−
*J23117
+
** 1µL SpeI
 +
** 1µL PstI
 +
** 2µL buffer 10x FastDigest
 +
** 6µL H2O
 +
 
 +
Second Digestion:
 +
* Biobricks:
 +
**K115017
 +
**I13504
 +
 
 +
* Mix:
 +
** 10µL of our plasmid with gene
 +
** 1µL XbaI
 +
** 1µL PstI
 +
** 2µL buffer 10x FastDigest
 +
 
 +
After this step, we separate the sequence we need from the sequence we don't with electrophoresis
  
−
10uL of our plasmid with gene
 
−
1uL XbaI
 
−
1uL PstI
 
−
2uL buffer 10x FD
 
−
After this step, we separate the sequence we need from the sequence we doesn't with electrophoresis
 
  
−
*K115017
 
−
*I13504
 
  
 
====Electrophoresis====
 
====Electrophoresis====
Line 45: Line 55:
 
''by Coralie, Audrey''
 
''by Coralie, Audrey''
  
−
2,5uL J23117
+
* J23117 + I13504
−
15uL I13504
+
**2,5µL J23117
−
1uL Ligase
+
**15µL I13504
−
2uL buffer 10x
+
**1µL T4 DNA Ligase
 +
**2µL Buffer T4 DNA Ligase 10x
  
−
1,5uL J23106
+
* J23106 + I13504
−
15,5uL I13504
+
**1,5µL J23106
−
1uL Ligase
+
**15,5µL I13504
−
2uL buffer 10x
+
**1µL TA DNA Ligase
 +
**2µL Buffer T4 DNA Ligase 10x
  
−
2uL J23101
+
* J23201 + I13504
−
14,5uL I13504
+
** 2µL J23101
−
1uL Ligase
+
** 14,5µL I13504
−
2uL buffer 10x
+
** 1µL Ligase
 +
** 2µL buffer 10x
  
−
Waiting all the night under 16°C
+
Incubation ON, 16°C
  
 
'''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

Revision as of 11:02, 17 July 2015

Wednesday 8th July

Lab Work

Transformation

by Audrey

Biobrick: R0051 (2015) On LB + Ampicillin 100ug/mL

Rehydratation

by Seong Koo, Johan

R0051 from different years (2013 and 2014)

Digestion

by Pauline, Coralie

First Digestion:

  • Biobricks:
    • J23101
    • J23106
    • J23117
  • Mix:
    • 10µL of our plasmid with promotor
    • 1µL SpeI
    • 1µL PstI
    • 2µL buffer 10x FastDigest
    • 6µL H2O

Second Digestion:

  • Biobricks:
    • K115017
    • I13504
  • Mix:
    • 10µL of our plasmid with gene
    • 1µL XbaI
    • 1µL PstI
    • 2µL buffer 10x FastDigest

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


Electrophoresis

by Johan, Coralie, Seong Koo, Pauline

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

  • K115017
  • I13504

Ligation

by Coralie, Audrey

  • J23117 + I13504
    • 2,5µL J23117
    • 15µL I13504
    • 1µL T4 DNA Ligase
    • 2µL Buffer T4 DNA Ligase 10x
  • J23106 + I13504
    • 1,5µL J23106
    • 15,5µL I13504
    • 1µL TA DNA Ligase
    • 2µL Buffer T4 DNA Ligase 10x
  • J23201 + I13504
    • 2µL J23101
    • 14,5µL I13504
    • 1µL Ligase
    • 2µL buffer 10x

Incubation ON, 16°C

Members present:

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