Figure 4. Schematic of making a template fragment of THSD1 from overlapping PCR.

Detailed information for the oligos is provided in Table 2. BamHI was added to the forward primer for NT-fragment PCR. Overlapping PCR contains part of a signal peptide (highlighted in the light blue box), BsaI DNA cassette (selected from Figure 2, highlighted in the red box), and part of THSD1 coding sequence (highlighted in the orange box). Bsu36I is in the middle of the reserve primer for CT fragment PCR. The detailed information of the final PCR product is indicated in the region of the template clone between two black lines.