Team:Paris Saclay/Notebook/August/6

From 2014.igem.org

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Then bain-marie at 37°C for one hour.
Then bain-marie at 37°C for one hour.
Finally conduct PCR cleanup for both BioBricks, with 15µl of NE buffer.
Finally conduct PCR cleanup for both BioBricks, with 15µl of NE buffer.
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====PCR pCola plasmide (Geranyl Synthase)====
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''by melanie''
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amplification of geranyl Synthase
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[https://2014.igem.org/Team:Paris_Saclay/Protocols/PCR_for_the_genomic_DNA_of_bacteria protocol]
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====Gel electrophoresis====
====Gel electrophoresis====

Revision as of 09:16, 7 August 2014

Contents

Wednesday 6th August

Lab work

A - The chassis coli Odor free

Final stock of MG1655

by Romain

2 final stocks of the MG1655 made with 1ml of strain and 250µl glycerol 87%, in -80°C.

C - Salicylate Inducible Suppressing System

Plasmid DNA Purification

by Eugene and Fabio

  • BBa_K1372000 Cl.1
  • BBa_K1372000 Cl.2

From Liquid Culture made the 5th August

Protocol

Digestion

by Eugene and Fabio

In order to place the BBa_B0015 just after the BBa_K1372000, we will start the [http://parts.igem.org/Help:Assembly/3A_Assembly Assembly procedure] again.

  • BioBrick BBa_K1372000 (Salicylate promoter + NahR + RNA suppressor) as Part A
  • BioBrick BBa_B0015 (Double terminator) as Part B

The enzymes used were:

  • EcoRI and SpeI for BBa_K1372000
  • EcoRI and XbaI for BBa_B0015

TODO: Illustration of the process

BioBrick Assembly - Digest Reaction Protocol

Segregate Process by Electrophoresis

by Eugene and Fabio

  1. BBa_K1372000 digested by EcoRI and SpeI
  2. BBa_B0015 digested by EcoRI and XbaI

Results:

In progress

BioBrick Assembly - Segregate Process Protocol

D - Lemon scent

PCR Cleanup

by Pierre & Sean

BioBricks used: BBa_K762100 and BBa_K517003 (PCR perfomed on the 4th August)

Protocol

Gel electrophoresis

by Melanie

Plasmid used: p cola

Results inconclusive. Wrong enzyme was used to prepare PCR.

PCR digestion

by Pierre & Sean


Protocol

For each BioBrick (Bba_K517003 and BBa_K762100)mix the following into a microcentrifuge tube.

component volume
H2O 5.5μl
PacI .5μl
Fast Digest buffer 4μl
Cleaned-up PCR content 30μl

Then bain-marie at 37°C for one hour. Finally conduct PCR cleanup for both BioBricks, with 15µl of NE buffer.

PCR pCola plasmide (Geranyl Synthase)

by melanie

amplification of geranyl Synthase

protocol



Gel electrophoresis

(with Melanie)

Samples used: BBa_K517003 (2µl), BBa_K762100 (2µl), p cola(12µl)

Bibliography

By Hoang Vu

Still working on my mission: find more informations about limonene, pinene and citral A and B.I got now a couple of very interessant reviews about limonene and pinene.


Members present:

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

Back to the calendar

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