Team:UT-Tokyo/Transformation

From 2010.igem.org

(Difference between revisions)
 
Line 17: Line 17:
to thaw out igem parts
to thaw out igem parts
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# With a pipette tip, punch a hole in the foil
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*1. With a pipette tip, punch a hole in the foil
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# Add 15uL of TE (MilliQ),and pipetting
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*2. Add 15uL of TE (MilliQ),and pipetting
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# Pipette 1uL of the resuspended DNA Transformation into your desired competent cells
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*3. Pipette 1uL of the resuspended DNA Transformation into your desired competent cells
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# Hold on ice for 30 mins
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*4. Hold on ice for 30 mins
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# Heat shock at 42°C for 45 seconds (and on ice after it)
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*5. Heat shock at 42°C for 45 seconds (and on ice after it)
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# Add 300uL of LBborth in each epp
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*6. Add 300uL of LBborth in each epp
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# Wait for 10 mins
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*7. Wait for 10 mins
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# Hold at 37℃ for 30 mins
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*8. Hold at 37℃ for 30 mins
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# Plate out
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*9. Plate out
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# Incubate at 37°C
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*10. Incubate at 37°C
{{UT-Tokyo_Foot}}
{{UT-Tokyo_Foot}}

Latest revision as of 07:05, 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