Team:Paris Saclay/Notebook/August/4

From 2014.igem.org

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Tubes were placed in PCR machine with the following parameters.
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Initial denaturation
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98°C
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1 min
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1
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Revision as of 14:00, 4 August 2014

Contents

Monday 4th August

Lab Work

A - The chassis coli Odor free

Striate on Dishes

by Juliette & Romain

We striated MG1655 and MG1655Z1 on LB and Kan dishes, from the striates prepared on LB dishes 1st August

C - Salicylate Inducible Suppressing System

Plasmid DNA Purification

by Eugene and Fabio

  • Ligation of BBa_J61051 and BBa_K228001 made the 30th July

From Liquid Culture made the 1st August

Digestion to check

In progress

Electrophoresis

In progress

D - Lemon scent

Gel electrophoresis of BBa_K762100

by Sean

Samples used were prepared on the 30th July.

Results

From left to right: 5 µl of BBa_K762100 + "old" Phusion enzyme, 5 µl of BBa_K762100 + "new" Phusion enzyme

Electroporation

by Terry

Strain used: DY330 containing plasmid pJBEI6409.

We had to switch the Limonen Synthase cassette with the Apramycin resistance.

The culture has been placed at 42°C for 17 min then cooled 10 min before centrifugation.

Polymerase chain reaction

by Sean & Pierre

BBa_K517003

component volume
H2O 35.5μl
Phusion buffer 5X 10μl
dNTPs 2μl
iPS68bis 1μl
iPS69 1μl
DNA 1μl
Phusion enzyme 0.5μl


BBa_K762100

For this BioBrick two tubes were prepared: one with undiluted DNA and one with DNA diluted 10-1.

component volume
H2O 35.5μl
Phusion buffer 5X 10μl
dNTPs 2μl
iPS66 1μl
iPS67 1μl
DNA 1μl
Phusion enzyme 0.5μl

Tubes were placed in PCR machine with the following parameters.

Cycle step Temperature Time Cycle

Initial denaturation

98°C

1 min

1

Denaturation 98°C 15 s 25 - 30
Annealing 52°C 25 s 25 - 30
Extension 72°C 45 s 25-30
Final extension 72°C 10 min 1
Final extension 8°C hold 1

Members there:

  • Instructors and advisors: Alice, Solenne and Sylvie.
  • Students: Eugene, Fabio, Hoang Vu, Juliette, Pierre, Romain, Sean and Terry.

Back to the calendar

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