Team:Paris Saclay/Notebook/August/7

Friday 7th August

Lab Work

Transformation

by Coralie

  • BBa_K1707032
  • BBa_K1707033
  • BBa_K1707026

As usual

Plasmid extraction

by Pauline

  • BBa_R0040 #1 and #2
  • BBa_K1707020 #1 and #2
  • BBa_K1707027 #1 and #2
  • BBa_K1707028 #1 and #2
  • Plasmid test #1 and #2

With Macherey-Nagel Extraction kit

Digestion verification

by Pauline

  • BBa_K1707020 #1 and #2
  • BBa_K1707027 #1 and #2
  • BBa_K1707028#1 and #2
  • Plasmid test #1 and #2

Mix:

  • 2 µL plasmid
  • 0,5 µL XbaI
  • 0,5 µL PstI
  • 1µL Buffer FastDigest 10x
  • 6 µL H2O

Incubation 1h30, 37°C

SpeI Test

by Pauline

Mix:

  • 2 µL BBa_K1707028 #1
  • 0,5 µL SpeI
  • 1µL Buffer N
  • 6,5 µL H2O

Incubation 2h30, 37°C

Electrophoresis

by Pauline

Agarose gel 1%, 100V

We can conlude that:

  • SpeI is OK
  • BBa_K1707020 #1 isn't OK
  • BBa_K1707020 #2 is OK
  • BBa_K1707027 #1 and #2 are OK
  • BBa_K1707028 #1 and #2: we have to check them another time
  • Plasmid test #1 and #2 are OK but not totally digested

New culture

by Pauline

  • BBa_R0051 #1

with 5mL LB and 5 µL Ampicillin


Digestion Verification

by Pauline

Mix:

  • 2 µL Plasmid
  • 0,5 µL XbaI
  • 0,5 µL PstI
  • 6 µL H2O
  • 1 µL Buffer FastDigest 10x


Low budget challenge

by Coralie We can see a lot of small colony. We put it ON 37°C.


Member present:

  • Instructors: Claire
  • Students: Coralie and Pauline

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