Huang: DH5alpha transformation: Difference between revisions
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(New page: 1. Set tubes of frozen cells on ice and let thaw very slowly 2. Add 100ul thawed cells to new eppendorf 3. Add 1-2ul DNA to cells 4. Incubate on ice for 20 min 5. Heat shock in 42C water b...) |
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1. Set tubes of frozen cells on ice and let thaw very slowly | ==Overview== | ||
2. Add 100ul thawed cells to new eppendorf | |||
3. Add 1-2ul DNA to cells | This protocol allows competent cell transformation. | ||
4. Incubate on ice for 20 min | |||
5. Heat shock in 42C water bath for one min (make sure water bath is at exactly 42C) | ==Materials== | ||
6. Incubate on ice 2 min | *DH5alpha competent cells | ||
7. Add 900ul growth medium (LB) and shake 1 hour at 37C | *LB | ||
8. Plate about 200ul and grow overnight | *LB + antibiotic plates | ||
*Plasmid DNA or DNA to transform the cells with | |||
==Procedure== | |||
*1. Set tubes of frozen cells on ice and let thaw very slowly | |||
*2. Add 100ul thawed cells to new eppendorf | |||
*3. Add 1-2ul DNA to cells | |||
*4. Incubate on ice for 20 min | |||
*5. Heat shock in 42C water bath for one min (make sure water bath is at exactly 42C) | |||
*6. Incubate on ice 2 min | |||
*7. Add 900ul growth medium (LB) and shake 1 hour at 37C | |||
*8. Plate about 200ul and grow overnight | |||
==Contact== | |||
*[[User:Carolina Tropini|Carolina Tropini]] | |||
Latest revision as of 16:49, 1 April 2009
Overview
This protocol allows competent cell transformation.
Materials
- DH5alpha competent cells
- LB
- LB + antibiotic plates
- Plasmid DNA or DNA to transform the cells with
Procedure
- 1. Set tubes of frozen cells on ice and let thaw very slowly
- 2. Add 100ul thawed cells to new eppendorf
- 3. Add 1-2ul DNA to cells
- 4. Incubate on ice for 20 min
- 5. Heat shock in 42C water bath for one min (make sure water bath is at exactly 42C)
- 6. Incubate on ice 2 min
- 7. Add 900ul growth medium (LB) and shake 1 hour at 37C
- 8. Plate about 200ul and grow overnight
