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並前往https://research.cbc.osu.edu/kudryashov.1/wp-content/uploads/2013/08/Site-Directed-Mutagenesis.pdf

Site Directed Mutagenesis Protocol

For calculating Tm for primers intended to introduce insertions or deletions, use this modified version of the above formula: Tm=81.5 + 0.41(%GC) - 675/N, where ...

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