Team:UT-Tokyo/Transformation

From 2010.igem.org

(Difference between revisions)
Line 15: Line 15:
<h2>Protocol</h2>
<h2>Protocol</h2>
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・to thaw out igem parts
+
to thaw out igem parts
-
1. With a pipette tip, punch a hole in the foil
+
 
-
2. Add 15uL of TE (MilliQ),and pipetting
+
# With a pipette tip, punch a hole in the foil
-
3. Pipette 1uL of the resuspended DNA Transformation into your desired competent cells
+
# Add 15uL of TE (MilliQ),and pipetting
-
4. Hold on ice for 30 mins
+
# Pipette 1uL of the resuspended DNA Transformation into your desired competent cells
-
5. Heat shock at 42°C for 45 seconds (and on ice after it)
+
# Hold on ice for 30 mins
-
6. Add 300uL of LBborth in each epp
+
# Heat shock at 42°C for 45 seconds (and on ice after it)
-
7. Wait for 10 mins
+
# Add 300uL of LBborth in each epp
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8. Hold at 37℃ for 30 mins
+
# Wait for 10 mins
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9. Plate out
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# Hold at 37℃ for 30 mins
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10. Incubate at 37°C
+
# Plate out
 +
# Incubate at 37°C
{{UT-Tokyo_Foot}}
{{UT-Tokyo_Foot}}

Revision as of 07:02, 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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