Theoligodesign for MAGE starts with the location of the target site in the replichore. To achieve the highest等位基因替换频率(ARF)when the oligo is annealed to the lagging strand, it is paramount to consider the strandedness during the oligo(s) design.
Therefore, we should design the oligo(s) sequence(s) depending on the target site location. You can have a look at the different areas in Figure 1.
Figure 1. Replichore sites.
Below, you can see the basic rules for oligo design, once the target site has been located in the replichore:
Other important parameters when designing the oligo(s) are:
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