User:Wilfred J. Poppinga/Notebook/Wilfreds Project/2009/08/25

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 * style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]] Project name
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


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Combining MBP-ArsR with inducible promotors

 * Cut plasmids containing promotor with SpeI and PstI
 * 1 μL 10x fast digest buffer
 * 0.5 μL SpeI
 * 0.5 μL PstI
 * 8 μL Vector
 * Incubate 1 h @ 37 °C
 * Purify cut plasmid using PCR clean up kit


 * Cut plasmids containing MBP-ArsR with XbaI and PstI
 * 1 μL 10x fast digest buffer
 * 0.5 μL XbaI
 * 0.5 μL PstI
 * 8 μL Vector
 * Incubate 1 h @ 37 °C
 * Put restriction on 1% Agarose (TBE) gel
 * Cut band of ~1500 bp out
 * Purify using gel purification kit
 * End volume 30 μL


 * Ligation
 * μL ligation buffer
 * 1 μL ligase
 * μL vector
 * μL insert
 * Incubate

to do

 * Watercampus
 * AquaExplorere
 * Wetsus
 * Rest
 * Safety!!