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IMC i02C Primer Design to Amplify Feature on Feature Lane

Perform PCR primer design to amplify selected features on the feature lane of the main feature map.

Operation

  1. Load a genomic sequence with features to PCR into the main feature map and make it the current sequence.
  2. Click the feature you want to amplify.
  3. Feature is selected and displayed in complementary color.
  4. Right-click on the feature on the feature.
  5. A pull-down menu will be displayed.
  6. Select PCR Primer Design.
  7. The PCR Primer Design dialog is displayed.
  8. Click Show Parameter ....
  9. The dialog will expand and the primer design parameters will be displayed.
  10. Select Outside if you want to amplify PCR products containing the entire selected features.
  11. Select Inside if you have amplified a portion of the inside of the selected feature.
  12. Click Start Design.
  13. The primer design is executed, and when finished, the Primer Design List dialog box will be displayed.
  14. All primer sets that satisfy the design conditions are displayed.
  15. Check on the top line.
  16. The region that the checked primer pair amplifies is displayed in color on the feature lane.
  17. Click Amplify.
  18. The confirmation message Start PCR? Is displayed.
  19. Click "Yes (Y)".
  20. The PCR reaction is executed and the Amplify List dialog box is displayed.
  21. Usually one product is displayed.
  22. I'll check it.
  23. Click Load.
  24. This PCR Product is loaded into the main current directory.
  25. A confirmation message Completed !!! is displayed.
  26. Click "OK".
  27. The confirmation message closes. Click the loaded PCR Product.
  28. The main feature map is switched and the feature map of that PCR Product is displayed.
  29. The amplified region contains the first selected feature.

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