How do I clone my gene of interest in the same translational reading frame as a tag present in the cloning vector (e.g., fluorescent protein, Myc, HA, etc.)?
How do I clone my gene of interest in the same translational reading frame as a tag present in the cloning vector (e.g., fluorescent protein, Myc, HA, etc.)?
Translational reading frame continuity with a tag is adjusted within the length of the gene-specific portion of the PCR primer, or by adding nucleotides between the gene-specific portion and the 15-nt homology of the PCR primer.
Please note that the current version of our online Primer Design Tool does not allow an adjustment for translational reading frame continuity. As such, the primer sequence should be manually designed by the user; we recommend SnapGene Viewer as a helpful, free online tool to help with this task.