Q1: What amount of the template DNA is added to the reaction mixture?

0.5-3 ng per 1 kbp DNA for 1 µL of the reaction mixture.
The amount of DNA is calculated based on number of DNA molecules. For example, in case of 6 kbp of plasmid, regardless of length of ORF, 3 to 18 ng of DNA is added for 1 µL of the reaction mixture.

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