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. Author manuscript; available in PMC: 2025 Dec 13.
Published in final edited form as: Nat Protoc. 2024 Dec 15;20(6):1546–1583. doi: 10.1038/s41596-024-01090-z
Component Amount per reaction (μL) Final concentration
NebNext High-Fidelity 2x PCR Master Mix (NEB) 12 1x
IVT Forward Primer (50uM) PD3771 (Table 1) 0.2 0.2μM
IVT Reverse Primer (50uM) PD3762 (Table 1) 0.2 0.2μM
Plasmid template (e.g. pDY0216) 10ng/μL 1 2ng/μL
UltraPure water To 50μL