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== Transformation of XL1-Blue Supercompetent Cells  ==

Notes Please read the Transformation Guidelines before proceeding with the transformation protocol. XL1-Blue cells are resistant to tetracycline. If the mutagenized plasmid contains only the tetR resistance marker, an alternative tetracycline-sensitive strain of competent cells must be used. 

1. Gently thaw the XL1-Blue supercompetent cells on ice. For each control and sample reaction to be transformed, aliquot 50 ÎŒl of the supercompetent cells to a prechilled 14-ml BD Falcon polypropylene round-bottom tube. 

2. Transfer 1 ÎŒl of the Dpn I-treated DNA from each control and sample reaction to separate aliquots of the supercompetent cells.<br>Note Carefully remove any residual mineral oil from the pipet tip before transferring the Dpn I-treated DNA to the transformation reaction. As an optional control, verify the transformation efficiency of the XL1-Blue supercompetent cells by adding 1 ÎŒl of the pUC18 control plasmid (0.1 ng/ÎŒl) to a 50-ÎŒl aliquot of the supercompetent cells. Swirl the transformation reactions gently to mix and incubate the reactions on ice for 30 minutes. 

3. Heat pulse the transformation reactions for 45 seconds at 42°C and then place the reactions on ice for 2 minutes.<br>Note This heat pulse has been optimized for transformation in 14-ml BD Falcon polypropylene round-bottom tubes.<br> 

[[Category:Protocols]]