Team:Paris Saclay/Notebook/August/14

From 2014.igem.org

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(Transformation of DH5α)
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[https://2014.igem.org/Team:Paris_Saclay/Protocols/Transformation_of_competent_e.coli_by_CaCl2 Protocol]
[https://2014.igem.org/Team:Paris_Saclay/Protocols/Transformation_of_competent_e.coli_by_CaCl2 Protocol]
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===D - Lemon Scent===
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===Lemon Scent===
''by Melanie''
''by Melanie''

Revision as of 09:04, 14 October 2014

Contents

Thursday 14th August

Lab Work

by Melanie

Preparation of DH5a CaCl2 Competent cell

Protocol

Lemon Scent

by Melanie

From the 20 ml of odor free culture (pJBEI Apra - pPSI) done yesterday (by Laetitia)

Plasmid extraction without Nucleospin protocol

+ Stock (DH5a pJBEI + apra)

Transformation of DH5α

by Sean & Eugene

(Cacl2 protocol)

We prepare a LB agar environment with a concentration of:

  • 0.5 mM of IPTG
  • 0.008% of Xgal
  • 1/2000 of Ampicilline

and we spread our preparation in a petri dish and incubate overnight at 37°C.

Human Practices

Art & Design

by Terry

We found an interesting colorless medium named M63.

Preparation of M63 medium, concentration 5X, one liter basis :

  • 10g (NH4)2 SO4
  • 68g KH2 PO4
  • 2.5mg FeSO4 7H2O
  • Adjust to pH 7 with KOH

The concentrated medium is diluted to 1X with sterile water and the following solution ( one liter basis ) :

  • 1 ml 1M MgSO4 7H2O
  • 10 ml 20% carbon source (sugar or glycerol)
  • 0.1 ml 0.5% vitamin B1

100 ml of M63 has been prepared for future manipulations.


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