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

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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
8μl buffer
2μl EcoRI
2μl XbaI
8μl MQ water
so 10μl mix with 10μl sample

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