Team:UT-Tokyo/Transformation

From 2010.igem.org

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<h1>Transformation</h1>
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<h2>Preparation</h2>
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*iGEM parts / ligation products
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*LBbroth (No antibiotic)　500μL
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*TE　15μL
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*plates
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*ice box
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*heat block(42℃)
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*competent cells
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→ always onice! Melt on ice! Mix DNA as soon as cells melt!
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<h2>Protocol</h2>
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・to thaw out igem parts
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1. With a pipette tip, punch a hole in the foil
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2. Add 15uL of TE (MilliQ),and pipetting
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3. Pipette 1uL of the resuspended DNA Transformation into your desired competent cells
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4. Hold on ice for 30 mins
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5. Heat shock at 42°C for 45 seconds (and on ice after it)
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6. Add 300uL of LBborth in each epp
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7. Wait for 10 mins
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8. Hold at 37℃ for 30 mins
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9. Plate out
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10. Incubate at 37°C
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{{UT-Tokyo_Foot}}

Revision as of 07:00, 23 September 2010

UT-Tokyo

Transformation

Preparation

  • iGEM parts / ligation products
  • LBbroth (No antibiotic)　500μL
  • TE　15μL
  • plates
  • ice box
  • heat block(42℃)
  • competent cells

→ always onice! Melt on ice! Mix DNA as soon as cells melt!

Protocol

・to thaw out igem parts 1. With a pipette tip, punch a hole in the foil 2. Add 15uL of TE (MilliQ),and pipetting 3. Pipette 1uL of the resuspended DNA Transformation into your desired competent cells 4. Hold on ice for 30 mins 5. Heat shock at 42°C for 45 seconds (and on ice after it) 6. Add 300uL of LBborth in each epp 7. Wait for 10 mins 8. Hold at 37℃ for 30 mins 9. Plate out 10. Incubate at 37°C


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