Generating a New sgRNA Vector, pGL3-U6-sgRNA-PGK-mRFP-T2A-PuroR, to Improve Base Editing: Table 3

Table 3: Touchdown PCR parameters for amplifying the mRFP-T2A insert.


Cycle	Denaturation	Annealing	Extension

1	94 °C, 5 min

2–7	94 °C, 30 sec	70 °C, 30 sec (1 °C less each cycle)	72 °C, 44 sec

8–31	94 °C, 30 sec	64 °C, 30 sec	72 °C, 44 sec

32			72 °C, 10 min