IGEM:Groningen/Notebook/iGEM 2011/2011/05/30

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30-5-11 Clone promotors upstream RBS-GFP-DT for eventual measurements of the promotor activities. Start working on strategy The following promotors are located on the following vectors: PBADaraC: pSB2K2 PcI: pSB2K2 PlasB: pSB1A1 Digest the promotor fragment with: EcoRI and SpeI Digest the vector with EcoRI and XbaI Cutmix: 8μl buffer 2μl EcoRI 2μl XbaI 8μl MQ water so 10μl mix with 10μl sample CutmixII: 8μl buffer 2μl EcoRI 2μl SpeI 8μl MQ water so 10μl mix with 10μl sample Ligation overnight, per sample: 1μl T4 DNA ligase 2μl T4 DNA ligase buffer 7μl insert 3μl plasmid 7μl MQ water