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Golden Gate Assembly of CRISPR sgRNAs into donor plasmids
| Components | Amount |
|---|---|
| Donor plasmid (U6 or H1 or 7SK) (100 ng/μL) | 1 μL |
| Annealed sgRNA oligonucleotides (1:200 diluted) | 2 μL |
| Tango buffer (10×) | 2 μL |
| DTT (10 mM) | 1 μL |
| ATP (10 mM) | 1 μL |
| FastDigest Esp3I restriction enzyme | 1 μL |
| T4 DNA ligase | 1 μL |
| ddH2O | 11 μL |
| Total | 20 μL |
