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Team:UT-Tokyo/Transformation
From 2010.igem.org
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<h2>Protocol</h2> | <h2>Protocol</h2> | ||
| - | + | to thaw out igem parts | |
| - | 1. With a pipette tip, punch a hole in the foil | + | |
| - | 2. Add 15uL of TE (MilliQ),and pipetting | + | *1. With a pipette tip, punch a hole in the foil |
| - | 3. Pipette 1uL of the resuspended DNA Transformation into your desired competent cells | + | *2. Add 15uL of TE (MilliQ),and pipetting |
| - | 4. Hold on ice for 30 mins | + | *3. 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) | + | *4. Hold on ice for 30 mins |
| - | 6. Add 300uL of LBborth in each epp | + | *5. Heat shock at 42°C for 45 seconds (and on ice after it) |
| - | 7. Wait for 10 mins | + | *6. Add 300uL of LBborth in each epp |
| - | 8. Hold at 37℃ for 30 mins | + | *7. Wait for 10 mins |
| - | 9. Plate out | + | *8. Hold at 37℃ for 30 mins |
| - | 10. Incubate at 37°C | + | *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
Copyright © 2010 iGEM UT-Tokyo. All rights reserved.
