Wednesday 12th August

Lab Work

Purification

by Pauline

BBa_I13602 #1
BBa_I13602 #2
BBa_K1707020
BBa_K1707030

With Macherey-Nagel Purification kit

Quantification

by Pauline

Agarose gel 1%, Migration 90V

Quantification, from left to right: 1. DNA Ladder, 2. BBa_K1707020, 3. BBa_K1707030, 4. BBa_I13602#1, 5. BBa_I13602#2, 6. Empty, 7. Empty, 8. Empty, 9. Empty, 10. Empty

We can conclude:

BBa_I13602 #1: 40 µg/µL
BBa_I13602 #2: 40 µg/µL
BBa_K1707020: 10 µg/µL
BBa_K1707030: 13 µg/µL

Digestion

by Coralie

BBa_K1707011 #3 and BBa_K1707012 #2
- Mix for each reaction:
  - 1 µL PstI
  - 1 µL XbaI
  - 2 µL Buffer FastDigest 10x
  - 6 µL H2O
  - 10 µL plasmid
- Incubation 37°C, 2h

Purification on gel

by Pauline

Agarose gel 1%, 90V

BBa_K1707011 #3
BBa_K1707012 #2

Verification of gel purification, from left to right: 1. DNA Ladder, 2. BBa_K1707011#3, 3. BBa_K1707012#2, 4. Empty, 5. Empty, 6. Empty, 7. Empty, 8. Empty, 9. Empty, 10. Empty

We can conclude that biobricks are OK

Purification by the Macherey-Nagel Purification kit

Ligation

by Pauline

BBa_K1707035: BBa_K1707030 + BBa_I13602
- 11,6 µL BBa_K1707030
- 1,25 µL BBa_R0040
- 2 µL Ligase
- 2 µL Buffer 10x
- 3,15 µL H2O

BBa_K1707036: BBa_K1707020 + BBa_I13602
- 15,5 µL BBa_K1707006
- 1,25 µL BBa_B0030
- 2 µL Ligase
- 2,5 µL Buffer 10x
- 3,75 µL H2O

Incubation 4°C, ON

New culture

by Pauline

From stock:
- BBa_K1707012 #2 x2
- BBa_K1707011 #3 x2
- BBa_R0051
Two clones from plate of each:
- BBa_K1707033
- BBa_K1707031
- BBa_K1707016

In 5mL LB + 5µL of appropriate antibiotic

Soil experiment

Member present:

Instructors: Claire
Students: Coralie and Pauline

Back to the calendar

Team:Paris Saclay/Notebook/August/12

Contents

Wednesday 12th August

Lab Work

Purification

Quantification

Digestion

Purification on gel

Ligation

New culture

Soil experiment