Team:Heidelberg/Notebook/Material Methods
From 2010.igem.org
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==Cell culture== | ==Cell culture== | ||
- | === | + | ===Media=== |
+ | Media: | ||
+ | *HeLa - DMEM, 10% FCS, Pen/Strep 1%, L-Glu 1% | ||
+ | *HEK - DMEM, 10% FCS, Pen/Strep 1%, L-Glu 1% | ||
+ | *HEK T-REx - DMEM, 10% FCS, Pen/Strep 1%, L-Glu 1%, blasticidin, zeocin | ||
+ | *HEK T-REx (with pcDNA5 integrated) - DMEM, 10% FCS, Pen/Strep 1%, L-Glu 1%, blasticidin, hygromycin | ||
+ | *Huh7 - DMEM, 10% FCS, Pen/Strep 1%, L-Glu 1%, 1% non essential amino acids | ||
+ | |||
+ | |||
+ | ===Passaging=== | ||
+ | *remove media | ||
+ | *wash cells one time in PBS (10ml) | ||
+ | *remove PBS; add 2 ml of trypsin-EDTA solution and incubate cells for 5 min at 37 °C | ||
+ | *add 5 ml of the according media | ||
+ | *take 1/10th of the cell suspension and plate out on the according dish (either p100 dish, 6 well plate or T-flask) | ||
+ | |||
+ | |||
+ | ===Coating=== | ||
+ | *add 15ul of poly-L-lysine solution to each well (make sure whole surface is covered with solution) | ||
+ | *leave for 30' in the incubator | ||
+ | *remove poly-L-lysine solution | ||
+ | *wash once with PBS | ||
+ | |||
{{:Team:Heidelberg/Pagemiddle}} | {{:Team:Heidelberg/Pagemiddle}} | ||
{{:Team:Heidelberg/Bottom}} | {{:Team:Heidelberg/Bottom}} |
Revision as of 09:41, 24 October 2010
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