16/09/13

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Revision as of 12:30, 16 September 2013 by Brettvl (Talk | contribs)

Contents

Digesting pSB1C3 backbone

  • Digesting for ligation with the already digested Tod operon PCR products
  • Following the iGEM official protocol
  • Using EcoRI-HF and PstI-HF
  • Exluding the DpnI from the master mix and adjusting the volume of water accordingly
  • Protocol can be found at: [http://parts.igem.org/Help:Protocols/Linearized_Plasmid_Backbones]

Digestion of the TOD genes PCR products

The PCR products of TODX, TODF and TOBG were digested with the enzymes EcoRI-HF and PstI-HF.

Protocol

SampleDNA volume (ul)cutsmart buffer (ul)EcoRI (ul)PstI (ul)dH2O (ul)
TODX8.720.50.58.3
TODF8.320.50.58.7
TOBG820.50.59
  • The samples were incubated for 1 hour at 37C.
  • Heat kill the enzymes at 80C for 20 minutes.

Running the backbone and RFP on gel

  • on 1% gel to check if the gel extraction worked.

Digestion of the pGEM plasmid extracted on the 13/09/13