Team:Paris Saclay/Notebook/August/14

Friday 14th August

Lab Work

Quantification

by Pauline

Biobricks:

• 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

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

Biobricks:

• BBa_K1707035
• BBa_K1707036

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

Plasmid extraction

by Pauline

BIobricks:

• BBa_K1707031 #3, #4, #5 and #6

With Macherey-Nagel Extraction kit

Digestion

by Pauline

Biobricks:

• 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_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 (2nd 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

Biobricks:

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

With Macherey-Nagel Purification kit

New culture

by Pauline

Biobrick: 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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