Team:Paris Saclay/Notebook/August/18

Tuesday 18th August

Lab Work

Digestion

by Pauline

Biobricks:

• BBa_K1707035 #1 and #2
• BBa_K1707036 #1 and #2

Mix for each plasmid:

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

Incubation 37°C, 3h

Plasmid extraction

by Pauline

BIobricks:

• BBa_K1707022 #1 and #2
• BBa_K1707023 #1 and #2
• BBa_K1707034 #1 and #2

With Sylvie Lautru's Protocol

Electrophoresis

by Pauline

Agarose gel 1%, migration 90V

Biobricks:

• BBa_K1707035 #1 and #2
• BBa_K1707036 #1 and #2

We can conclude that BBa_K1707035 #1, BBa_K1707036 #1 and #2 are OK, but not BBa_K1707035 #2

Inoculation

by Pauline

3 strains: 1320; 1693; 1696

in 10mL LB without antibiotic

PCR

by Audrey

Biobricks:

• BBa_K1707000 #2, #3, #4
• BBa_K1707013 #1
• BBa_K1707030 #1
• BBa_K1707019 #1
• BBa_K1707020 #2
• BBa_K1707035 #1
• BBa_K1707036 #1
• BBa_K1707027 #1
• BBa_K1707021 #2

Control +: BBa_K115017 Control -: BBA_J23101+GFP (Interlab study)

Vtot in each tube= 50µL: 2µL plasmid + 48µL mix

Mix for all tubs:

• 476,25 µL H2O
• 150 µL Buffer 5X
• 7,5 µL Forward Primer (iPS43)
• 7,5 µL Reverse Primer (iPS3)
• 15 µL dNTP
• 3,75 µL GoTAQ
• 60 µL MgCl2

Digestion

by Pauline

Biobricks:

• BBa_K1707022 #1 and #2
• BBa_K1707023 #1 and #2
• BBa_K1707034 #1 and #2

Mix for each plasmid:

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

Incubation 37°C, 1h

Transplant

by Pauline

Biobrick: BBa_K1707031 clone #7 to #26 In a liquid culture: 5mL LB + 5µL Antibiotic

PCR

by Audrey

Member present:

• Instructors: Claire
• Students: Pauline and Audrey

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