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Monday 27th July

Lab Work

New culture

by Seong Koo

Biobricks:

• BBa_K1707005
• BBa_K1707006
• BBa_K1707007

Ligation

by Pauline and Coralie

• BBa_K1707008
  • 2µL Tp 10x
  • 1µL Ligase
  • 11µL BBa_K1707003
  • 6µL BBa_J23101

• BBa_K1707010
  • 1µL Tp 10x
  • 1µL Ligase
  • 12µL BBa_K1399005
  • 10µL BBa_B0015
  • 1µL H20

• BBa_K1707009
  • 2µL Tp 10x
  • 2µL Ligase
  • 10µL BBa_S03518
  • 6µL BBa_R0051

• BBa_K1707004
  • 3µL Tp 10x
  • 2µL Ligase
  • 8µL BBa_K1707003
  • 7µL BBa_R0051

Transformation

by Pauline and Coralie

Biobricks:

• BBa_K1707008
• BBa_K1707010
• BBa_K1707009
• BBa_K1707004

Digestion + Purification + Quantification

by Audrey

Biobricks:

• BBa_R0051
• BBa_K1707000

Digestion: as usual, with SpeI + PstI

Migration: Agarose gel 1%, migration 100V

Plasmid extraction

by Pauline

Biobricks:

• BBa_K1707003 #4 et #5

With Plasmid DNA purification kit from Macherey-Nagel

Digestion + Purification on agarose gel

by Pauline

Biobricks:

• BBa_K1707003 #4 and #5

Mix:

• 10µL plasmid BBa_K1707003
• 1µL XbaI
• 1µL PstI
• 2µL Buffer FastDigest
• 6µL H2O

Purification on agarose gel 1%, migration 100V

Wells 2-3: Quantification, Wells 9-10: Verification of purification; from left to right: 1. DNA Ladder, 2. BBa_K1707000#2, 3. BBa_50051, 4. Empty, 5. Empty, 6. Empty, 7. Empty, 8. Empty, 9. BBa_K1707003#1, 10. BBa_K1707003#2, 11. Empty 12. Empty

Cut with a scalpel Purification with the PCR Clean up kit from Macherey-Nagel

New Culture

by Johan

Strains:

• 1696
• 1693
• 1320

in 150mL LB without any antibiotic

Member present:

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

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