Team:Paris Saclay/Notebook/August/14

Friday 14th August

Lab Work

Quantification

by Pauline

  • BBa_R0051 (2nd send)
  • BBa_K1707011 #3
  • BBa_K1707012 #2
  • BBa_K1707033 #1 and #2

Mix:

  • 2 µL plasmid
  • 2 µL DNA Loading 6X
  • 6 µL H2O

Agarose gel 1%, Migration 90V

ParisSaclay 14.08.15-quantif(3).jpg

Quantification, from left to right: 1. DNA Ladder, 2. BBa_K1707011#3 (08/12), 3. BBa_K1707011#3 (08/12), 4. BBa_K1707012#2 (08/12), 5. BBa_K1707012#2 (08/13), 6. BBa_K1707033#1, 7. BBa_K1707033#2, 8. BBa_R0051 (2nd send), 9. Empty, 10. Empty

We can conclude:

  • BBa_R0051 (2nd send): 13 µg/µL
  • BBa_K1707011 #3: 40 µg/µL
  • BBa_K1707012 #2: 35 µg/µL
  • BBa_K1707033 #1 and #2: 20 µg/µL


New culture

by Pauline

  • BBa_K1707035
  • BBa_K1707036

We put 2 new clones in 5mL LB + 5µL Chloramphenicol


Plasmid extraction

by Pauline

  • BBa_K1707031 #3, #4, #5 and #6

With Macherey-Nagel Extraction kit


Digestion

by Pauline

  • BBa_K1707031 #3, #4, #5 and #6

Mix for each:

  • BBa_K1707011 and BBa_K1707012
    • 0,5 µL XbaI
    • 0,5 µL PstI
    • 1 µL FastDigest Buffer 10x
    • 6 µL H2O
    • 2 µL plasmid

Incubation 37°C, 1H30

Ligation

by Pauline

  • BBa_K1707023: BBa_K1707012 and BBa_J23101
    • 5,9 µL BBa_K1707012
    • 5 µL BBa_J23101
    • 2 µL Ligase
    • 1,5 µL Buffer
    • 0,6 µL H2O
  • BBa_K1707022: BBa_K1707011 and BBa_R0051
    • 5,4 µL BBa_K1707011
    • 7,6 µL BBa_R0051
    • 2 µL Ligase
    • 2 µL Buffer
    • 3 µL H2O
  • BBa_K1707034: BBa_K1707033 and BBa_J23101
    • 11 µL BBa_K1707033
    • 5 µL BBa_J23101
    • 2 µL Ligase
    • 2 µL Buffer

Incubation 3h, 4°C

Electrophoresis

by Pauline

Agarose gel, 1%, Migration 90V

ParisSaclay 14.08.15-vérif.jpg

Verification of gel purification, from left to right: 1. DNA Ladder, 2. BBa_K1707030#3, 3. BBa_K1707031#4, 4. BBa_K1707031#5, 5. BBa_K1707031#6, 6. Empty, 7. Empty, 8. Empty, 9. Empty, 10. Empty

Biobricks:

  • Verification:
    • BBa_K1707031 #3, #4, #5 and #6

We can conclude that all clones aren't OK because the band viewed corresponding to the BBa_R0040 plasmid

Transformation

by Pauline

  • BBa_K1707022
  • BBa_K1707023
  • BBa_K1707034

Soil experiment

by Coralie and Johan

Low budget experiment

by Coralie

Member present:

  • Instructors: Claire
  • Students: Coralie and Pauline

Back to the calendar

Contact
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t@iGEMParisSaclay
aUniversité Paris Sud
a91405 Orsay, France Copyright © 2015 iGEM Paris Saclay. All rights reserved All inspirations encouraged.
*This wiki was conceived during long nights of work and a great coffee experience in the beautiful Orsay, France.