Team:Paris Saclay/Notebook/July/22

Wednesday 22nd July

Lab Work

Electrophoresis

by Johan and Pauline

  • PCR Products:
    • BBa_J23101
    • BBa_K1707000
  • Purification quantification:
    • BBa_K1707002
    • BBa_J23101
    • BBa_R0051
    • BBa_K098997
    • BBa_K1707001
  • Undigested plasmids:
    • BBa_K1707001 #1 and #2
    • BBa_K1707002 #1 and #2
    • BBa_K1707003 #2
    • BBa_J23101
    • BBa_B0015

We can't see PCR products. We can suppose that they run out the gel.

Paris Saclay-22.07.2015 - Plasmides 2.jpg

Quantification, from left to right: 1. DNA Ladder, 2. BBa_K1707001#1, 3. BBa_K1707001#2, 4. BBa_K1707002#1, 5. BBa_K1707002#2, 6. BBa_K1707003#2, 7. BBa_J23101, 8. BBa_B0015, 9. Empty, 10. Empty, 11. Empty, 12. Empty

Paris Saclay-22.07.15 - Quantification 1.jpg

Quantification, from left to right: 1. DNA Ladder, 2. PCR J23101, 3. PCR K1707000#1, 4. PCR K1707000#2, 5. BBa_K1707002, 6. BBa_J23101, 7. BBa_R0051, 8. BBa_K098997, 9. BBa_K1707001, 10. Empty

Digestion

by Coralie

  • Digestion by EcoRI
    • Mix:
      • 7µL EcoRI
      • 7µL Buffer FastDigest 10x
      • 42µL H2O

8µL in each tube + 2µL Plamid

    • Biobricks:
      • BBa_B0015
      • BBa_J23101
      • BBa_K1707003 #2
      • BBa_K1707002 #1 and #2
      • BBa_K1707001 #1 and #2
  • Digestion by EcoRI + PstI
    • Mix:
      • 7µL EcoRI
      • 7µL PstI
      • 7µL Buffer FastDigest 10x
      • 35µL H2O

8µL in each tube + 2µL Plamid

    • Biobricks:
      • BBa_B0015
      • BBa_J23101
      • BBa_K1707003 #2
      • BBa_K1707002 #1 and #2
      • BBa_K1707001 #1 and #2

Incubation 2h, 37°C

Electrophoresis

by Johan

Paris Saclay-22.07.2015 - Gel vérif simple digestion EcoRI.jpg

Verification by digestion with EcoRI, from left to right: 1. DNA Ladder, 2. BBa_B0015, 3. BBa_J23101, 4. BBa_K1707003#2, 5. BBa_K1707002#2, 6. BBa_K1707002#1, 7. BBa_K1707001#2, 8. BBa_K1707001#1, 9. Empty, 10. Empty

Paris Saclay-22.07.15 - Vérification double digestion EcoRI + PstI.jpg

Verification by digestion with EcoRI and PstI, from left to right: 1. DNA Ladder, 2. BBa_B0015, 3. BBa_J23101, 4. BBa_K1707003#2, 5. BBa_K1707002#2, 6. BBa_K1707002#1, 7. BBa_K1707001#2, 8. BBa_K1707001#1, 9. Empty, 10. Empty, 11. Empty, 12. Empty

We can conclude that K1707003 #2 isn't OK

Plasmid extraction

by Pauline and Seong Koo

Biobricks:

  • BBa_K1707003 #4 to #9
  • BBa_C0051
  • BBa_E0240
  • BBa_J06702


Soil experiment

by Audrey and Johan

Plates observation and count of CFU:

  • Strain 1696:
    • Dilution 1: more than 1000
    • Dilution 0.1: more than 1000
    • Dilution 0.01: 441 CFU
  • Strain 1693:
    • Dilution 1: more than 1000
    • Dilution 0.1: more than 1000
    • Dilution 0.01: 414 CFU
  • Strain 1320:
    • Dilution 1: more than 1000
    • Dilution 0.1: 1744 CFU
    • Dilution 0.01: 272 CFU

We take 1g of contaminated soil and treat it like yesterday to spread it on specific plates Incubation ON, 37°C

Electrophoresis of PCR products

ParisSaclay 22.07.15 ARN tige boucle PCR.jpg

Verification PCR products, from left to right: 1. DNA Ladder, 2. Indicator (BBa_J23101), 3. BBa_K1707000#1, 4. BBa_K1707000#2

Member present:

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

Back to the calendar

Contact
eigem.parissaclay AT gmail DOT com
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.