Team:Paris Saclay/Notebook/August/6

From 2014.igem.org

Contents

Wednesday 6th August

Lab work

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.

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

Paris Saclay 060814 cut gel.jpg

by Eugene and Fabio

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


BioBrick Assembly - Segregate Process Protocol

Lemon scent

PCR Clean-up

by Pierre & Sean

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

Protocol

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.

We acidentally used PvuI instead of PacI. - SC 6/8/14

PCR pCola plasmide (Geranyl Synthase)

by melanie

amplification of geranyl Synthase

protocol

PCR protocol: 98° --> 2min

5 PCR cycle:

time 10sec 20sec 45sec
temperature 98° (depending on the tube - see the gradient) 72°

25 PCR cycle

time 10sec 20sec 45sec
temperature 98° 72° 72°

and last step: 72° during 10min

but we don't have any results

Gel electrophoresis

Sean&Pierre&Melanie.JPG

by Sean,Pierre & Melanie

Samples used:

  1. BBa_K762100 (2µl)
  2. BBa_K517003 (2µl)
  3. 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.

Photo of the Day

Paris Saclay 6 august.jpg

Members present:

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

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