User:Wilfred J. Poppinga/Notebook/Wilfreds Project/2009/09/09

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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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Competent cells: TOP10 & DB3.1

 * 10 mL ON culture is used to inoculate 2·10 mL of LB, 50 μL per 10 mL.
 * Cultures are grown @ 37 °C untill an OD600 of 0.2 ~ 0.3 is reached.
 * Cultures are spinned down 5' @ 4000 rpm, 4 °C
 * Supernatant is removed and pellet is resuspended in 5 mL chilled 0.1 M CaCl2
 * Suspension is incubated on ice for 10 min.
 * Suspensions are spinned down 5' @ 4000 rpm, 4 °C
 * Supernatant is removed and pellet is resuspended in 2000 μL chilled 0.1 M CaCl2+10% glycerol prior to making aliquots.
 * Cells are divided in 50 μL aliquotes
 * Cells are stored @ -80 °C
 * Cells are stored @ -80 °C

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{| width="800"
 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] Project name
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


 * colspan="2"|
 * colspan="2"|

Entry title

 * Insert content here...

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