Team:Groningen/Protocols

From 2010.igem.org

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[[CaCl2 Competent Cells]]
[[CaCl2 Competent Cells]]
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'''Day 1:'''
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[[Growing Streptomyces]]
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-Inoculate 5 ml [[LB( TY)  medium]] with ''E.coli'' from glycerol stock
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-Grow overnight at 37 oC
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'''Day 2:'''
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-Inoculate 20 ml LB medium with 200 μl overnight culture
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-Grow at 37 oC until OD600 = 0,3-0,5(+/- 2 hours)
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-Spin down 5 minute at 4000 rpm at 4 oC
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-Resuspend in 10 ml chilled 0,1 M CaCl2 (from here, keep on ice! )
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-Inoculate on ice for 20 minutes
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-Spin down as before
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-Remove supernatant
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-Resuspend in 2 ml 0,1 M CaCl2 /10% glycerol
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-Divide 100 μl aliquots
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-Store competent cells in - 80 oC
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Revision as of 11:31, 24 August 2010

CaCl2 Competent Cells

Growing Streptomyces