How can I prepare primer (for mutagenesis), which will contain stop codon in first reading frame and will continue in second reading frame?

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Uwe Borgmeyer

If I understand you right, you have to choose a primer starting with a stop codon in frame 1: tca 123 456 ...

If this is your alternative reading frame: xxt ca1 234 567 .... you may choose ac for xx and thereby introduce an additional serine.

In other words, if your stop codon is not there in the natural situation your second protein will have depending onyour surrounding sequence 1 to 3 additional amino acids.

Alžběta Dostálková

Thank you very much for your responding! It is helpful.