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From 2012.igem.org

Mutagenesis

QuikChange® Lightning Site-Directed Mutagenesis Kit

Prepare the sample reaction(s) as indicated below

1. 5 μl of 10× reaction buffer
2. X μl (10–100 ng) of dsDNA template
3. X μl (125 ng) of oligonucleotide primer #1
4. X μl (125 ng) of oligonucleotide primer #2
5. 1 μl of dNTP mix
6. 1.5 μl of QuikSolution reagent
7. ddH2O to a final volume of 50 μl
8. Then add: 1 μl of QuikChange® Lightning Enzyme

Cycling Parameters for the QuikChange Lightning Site-Directed Mutagenesis Method

* For example, a 5-kb plasmid requires 2.5 minutes per cycle at 68°C.