Wisconsin/Protocol:Transformation
From 2007.igem.org
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#Heat shock in 42<sup>o</sup>C water bath for 30 seconds | #Heat shock in 42<sup>o</sup>C water bath for 30 seconds | ||
#Incubate on ice for 2 minutes | #Incubate on ice for 2 minutes | ||
| - | # | + | #Transfer cells to new culture tubes (more O<sub>2</sub> for growth) and add 900uL of SOC to each culture tube |
#Incubate on shaker at 37<sup>o</sup>C for 1 hour | #Incubate on shaker at 37<sup>o</sup>C for 1 hour | ||
#Spread ??? uL onto plate... centrifuge first??? | #Spread ??? uL onto plate... centrifuge first??? | ||
#Incubate plates for 16 hours at 37<sup>o</sup>C. Put plates upside down to prevent condensation on surface. | #Incubate plates for 16 hours at 37<sup>o</sup>C. Put plates upside down to prevent condensation on surface. | ||
Revision as of 22:41, 5 July 2007
Transforming Chemically Competent Cells
- Thaw cells on ice
- Pippet 100uL of cells into three 1.5mL tube. One for transformation and the other two for control.
- Normal: pippet 9uL of DNA into tube
- Positive Control: pippet 2uL of pUC18 or pUC19 into tube
- Negative Control: nothing
- Sit on ice for 30 minutes
- Heat shock in 42oC water bath for 30 seconds
- Incubate on ice for 2 minutes
- Transfer cells to new culture tubes (more O2 for growth) and add 900uL of SOC to each culture tube
- Incubate on shaker at 37oC for 1 hour
- Spread ??? uL onto plate... centrifuge first???
- Incubate plates for 16 hours at 37oC. Put plates upside down to prevent condensation on surface.
