Annealing Protocol
- Phosphorylation of the oligos
- 5.6μL DNAse/RNAse free water
- 6.0μL o15.011 (10µM)
- 6.0μL o15.012 (10µM)
- 2.0μL 10X T4 DNA ligase buffer
- 0.4μL T4 PolyNucleotide Kinase
Total: 20μL
- incube 30min at 37°C
- add 1μL of 1M NaCl
- incube 5min at 95°C
- let the mix cool down
- use 2μL of the mix as a 10X solution
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