Team:CU-Boulder/Notebook/Experiments

From 2013.igem.org

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[https://2013.igem.org/Team:CU-Boulder/Notebook/Experiments/Experiment 1.1 - Chemical Transformation Efficiency]
[https://2013.igem.org/Team:CU-Boulder/Notebook/Experiments/Experiment 1.1 - Chemical Transformation Efficiency]
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Just testing this right now...
 
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Experiment 1.1 – Chemical Transformation Efficiency
 
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Purpose:
 
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To explore the effects of competent cell : DNA ratio and heat shock temperature and duration on chemical transformation efficiency.
 
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Setup:
 
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Trial V (μL) cells V (μL) pUC19 Heat Shock Resistance
 
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1 (2x) 50 0 - -
 
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2 (2x) 170 1 10 min @ 37° C Ampicillin
 
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3 (2x) 50 2 10 min @ 37° C Ampicillin
 
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4 (2x) 50 2 30 s @ 42° C Ampicillin
 
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5 (2x) 50 2 60 s @ 42° C Ampicillin
 
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Procedure:
 
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1. Thaw competent cells on ice for ~15 minutes
 
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2. Aliquot 50 mL competent cells into 1.5 mL Eppendorf tubes
 
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3. Add pUC19 into competent cell aliquots
 
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4. Incubate on ice for ~30 minutes
 
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5. Use water bath to heat shock samples at various conditions
 
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6. Add 1 mL SOC recovery medium to each sample
 
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7. Allow cells to recover in shaker @ 37° C for ~1 hr.
 
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8. Plate 200 μL of each sample and allow to dry
 
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9. Store plates upside down in incubator @ 37° C overnight
 
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Results:
 
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Discussion:
 

Revision as of 17:22, 14 August 2013

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1.1 - Chemical Transformation Efficiency

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