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. 2017 Dec 5;7(23):e2625. doi: 10.21769/BioProtoc.2625

Table 7. Reagents for L1 Golden Gate assembly.

Reagent Quantity
pICH47732 _TpFCP:NAT 40 fmol
pICH47742_TpFCP:Cas9:YFP 40 fmol
pICH47751_TpU6:sgRNA1 40 fmol
pICH47761_TpU6:sgRNA2 40 fmol
pICH41780 (L4E linker) 40 fmol
pAGM4723 (L2 backbone) 40 fmol
BpiI 10 U
T4 DNA ligase 10 U
10x ligation buffer 2 µl
Nuclease-free water Up to 20 µl