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

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

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

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

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

Back to the calendar

Team:Paris Saclay/Notebook/August/18

Contents

Tuesday 18th August

Lab Work

Digestion

Plasmid extraction

Electrophoresis

Inoculation

PCR

Digestion

Transplant

PCR