- Change the selection to 'Mutagenesis' in the "Select Project Type" box in the left column. ('Cloning' will be selected by default upon opening the primer design tool.)
Deleting a specific sequence from any vector
The following protocol walks you through the steps for designing PCR primers to delete a sequence from any vector with In-Fusion Cloning. Use our online primer design tool to:
- add the vector sequence of your choice,
- specify the exact nucleotides to be deleted, and
- download primer and PCR information based on your design.
A. Select project type
B. Input vector sequence
Use the "Add Vector To Mutagenize" box to input your vector sequence in one of the following ways:
Provide your own vector sequence
- Click on 'Input sequence'.
- Paste your nucleotide sequence into the text box as plain text, without nucleotide or line numbering.
Choose from the preloaded list of vectors
- Click on the plus sign next to "Select a Takara or Clontech Vector".
- Choose a vector from the list.
C. Select sequence to delete
- In the pane on the right side of the screen, navigate to the area you want to modify by clicking on the vector map image to jump to a general area or feature.
- Use the nucleotide view to select and highlight the exact sequence you wish to delete.
D. Delete selected sequence
- In the "Select Modification" box, click the [Delete Selection] button.
- Double-check that your construct map and sequence show the correct region marked for deletion.
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