Team:Paris Saclay/Notebook/August/19

Wednesday 19th August

Lab Work

Electrophoresis

by Pauline

Agarose gel 1%, Migration 90V Biobricks:

• BBa_K1707000 #2, #3, #4
• BBa_K1707013 #1
• BBa_K1707030 #1
• BBa_K1707019 #1
• BBa_K1707020 #2
• BBa_K1707021 #2
• BBa_K1707035 #1
• BBa_K1707036 #1
• BBa_K1707027 #1
• pIJ773
• BBa_K1707022 #1 and #2
• BBa_K1707023 #1 and #2
• BBa_K1707034 #1 and #2

Digestion

by Pauline

• BBa_K1707022 #1 and BBa_K1707028 #1
  • 10 µL plasmid
  • 1 µL XbaI
  • 1 µL PstI
  • 2 µL Buffer FastDigest 10x
  • 6 µL H2O

• BBa_K1707034 #1
  • 10 µL plasmid
  • 1 µL SpeI
  • 2 µL PstI
  • 2 µL Tango Buffer 10x
  • 5 µL H2O

• BBa_K1707023 #1 and #2
  • 5 µL plasmid
  • 0,5 µL XbaI
  • 0,5 µL PstI
  • 1 µL Buffer FastDigest 10x
  • 3 µL H2O

Incubation 37°C, 1h30

Electrophoresis

by Pauline

Biobricks:

• BBa_K1707023 #1 and #2
• BBa_K1707022 #1
• BBa_K1707028 #1

Agarose gel 1%, 90V

We can conclude that BBa_K1707023 #1 and #2 are OK. We can cut BBa_K170708 #1 and BBa_K1707022 #1 bands.

PCR

by Pauline

Biobricks:

• BBa_K1707031 #7 to #26

We put 30 µL H2O in eqch tub. We add then the plasmid. And after, the mix.

Mix for each tub:

• 10 µL Buffer
• 0,25 µL Forward primer (iPS43)
• 0,25 µL Reverse primer (iPS44)
• 1 µL dNTP
• 0,25 µL GoTAQ
• 4 µL MgCl2
• 4,25 µL H2O

Digestion

by Pauline and Audrey

Biobricks: BBa_K1707004 #2

Mix:

• 15 µL plasmid
• 3 µL XbaI
• 1,5 µL SpeI
• 3 µL Tango Buffer 10x
• 7,5 µL H2O

Biobrick: BBa_K1707023 #1

Mix:

• 20 µL plasmid
• 2 µL PstI
• 1 µL SpeI
• 3 µL Tango Buffer 10x
• 4 µL H2O

Incubation 37°C, ON

Member present:

• Instructors: Claire
• Students: Pauline and Audrey

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