Protocol/2

From 2010.igem.org

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Protocol 2: 5 mL Overnight
 
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What you will need:
 
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* 10 mL culture tube Use 16mm x 160mm or 16mm x 125 mm
 
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* 5 mL LB
 
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* concentrated antibiotics
 
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* Single colonies on a plate (Best not to start O/N from glycerol stocks)
 
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Procedure:
 
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* Pipet 1.5-20uL of the appropriate antibiotic to the appropriate working concentration into culture tube.
 
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* Add 5 mL non-contaminated LB. Do this first, then add antibiotic
 
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* Select single colony using a sterile instrument that has been cooled and inoculate the liquid culture.
 
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* Place culture tube in incubator at 37 C overnight shaking vigorously (250 rpm).
 
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Notes:
 
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For I0500, culture overnight; then add 1 mM ITPG in morning and continue culture for another 3 hrs. Then miniprep.
 
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[[Team:Alberta/Notebook/protocols| Back]]
 
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Latest revision as of 17:16, 26 October 2010