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Thursday 13th August

Lab Work

Plasmid extraction

by Pauline

• BBa_R0051 (2nd send)
• BBa_K1707011 #3 (x2)
• BBa_K1707012 #2 (x2)
• BBa_K1707033 #1 and #2
• BBa_K1707031 #1 and #2
• BBa_K1707016 #1 and #2

With Macherey-Nagel Extraction kit (BBa_K1707011 #1 and #2 are assembled in the purification column) (BBa_K1707012 #1 and #2 are assembled in the purification column)

Digestion

by Coralie

• BBa_K1707011
• BBa_K1707012
• BBa_K1707016 #1 and #2
• BBa_K1707031 #1 and #2
• BBa_K1707033 #1 and #2
• BBa_R0051

Mixes:

• BBa_K1707011 and BBa_K1707012
  • 1 µL XbaI
  • 2 µL PstI
  • 2 µL FastDigest Buffer 10x
  • 1 µL H2O
  • 15 µL plasmid

• BBa_K1707016 #1 and #2 and BBa_K1707031 #1 and #2
  • 1 µL XbaI
  • 2 µL PstI
  • 2 µL FastDigest Buffer 10x
  • 11 µL H2O
  • 5 µL plasmid

• BBa_K1707033 #1 and #2
  • 1 µL XbaI
  • 2 µL PstI
  • 2 µL FastDigest Buffer 10x
  • 6 µL H2O
  • 10 µL plasmid

• BBa_R0051
  • 1 µL SpeI
  • 2 µL PstI
  • 2 µL Tango Buffer 10x
  • 5 µL H2O
  • 10 µL plasmid

Incubation 1h30, 37°C

Electrophoresis

by Pauline

Agarose gel, 1%, Migration 90V

Biobricks:

• Verification:
  • BBa_K1707016 #1 and #2
  • BBa_K1707031 #1 and #2
• Purification:
  • BBa_K1707011 #3
  • BBa_K1707012 #2
  • BBa_K1707033 #1 and #2
  • BBa_R0051 (2^nd send)

Wells 2-5: Verification by digestion with XbaI and PstI, Wells 6-10: Verification of gel purification; from left to right: 1. DNA Ladder, 2. BBa_K1707016#1, 3. BBa_K1707016#2, 4. BBa_K1707031#1, 5. BBa_K1707031#2, 6. BBa_K1707033#1, 7. BBa_K1707033#2, 8. BBa_K1707011#3, 9. BBa_K1707012#2, 10. BBa_R0051 (2^nd send)

We can cut all purification bands. For the verification, we can say that BBa_K1707016 #1 and #2 are OK, but not BBa_K1707031 #1 or #2.

We made a stock of BBa_K1707016 #1 and #2 and BBa_K1707033 #1 and #2.

Purification

by Pauline

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

With Macherey-Nagel Purification kit

New culture

by Pauline

• BBa_K1707031

We put 4 new clones in 5mL LB + 5µL Chloramphenicol

Soil experiment

by Coralie, Ibtissam, Johan and Claire

Member present:

• Instructors: Claire
• Students: Coralie and Pauline

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