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 | ||
- | + | *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 | |
{{UT-Tokyo_Foot}} | {{UT-Tokyo_Foot}} |
Latest revision as of 07:05, 23 September 2010
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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