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Table 3 TPCR conditions for cloning both gRNA into pCFD4d plasmid. The denaturation-annealing-elongation cycle was repeated 30 times

From: Efficient In Vivo Introduction of Point Mutations Using ssODN and a Co-CRISPR Approach

Temperature (°C)

duration

 

98

30 s

Initial denaturation

98

10 s

Denaturation

61

1 min

Annealing

72

7 min

Elongation

72

7 min

Final extension

10

Storage