View source for Team:Paris Saclay/Notebook/August/18

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=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
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