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IMC i01L Linearize Circular Vector by Restriction Enzyme

Linearize circular vector with restriction enzyme

Operation

  1. Load the DNA sequence of a circular vector into the current main directory.
  2. Load the sample sequence "pColdV.gbk" as an example.
  3. The sequence loaded in the main feature map is displayed.
  4. Find a restriction enzyme that cleaves this vector in only one place.
  5. Click the "Cloning -> Restriction Enzyme -> RE Registration & Editing ..." button from the menu.
  6. The "Enzyme Selection" dialog is displayed.
  7. In the "Select Enzyme" section, in the "Result" field, check "Any Enzyme".
  8. Enter 1 in the "which have from" and "To" fields.
  9. Click "Select All". All restriction enzymes are checked.
  10. Click "Show Recognition Site ...".
  11. The "Recognition Site" dialog is displayed.
  12. For example, you know that BamHI disconnects MCS in only one place, so check BamHI.
  13. The recognition site is displayed in the right pane.
  14. Check the BamHI in the right pane and click the "Digestion" button.
  15. A confirmation message will be displayed.
  16. Click "Yes (Y)".
  17. The "Digestion list" window will be displayed.
  18. Only one digested fragment is listed.
  19. Check the fragment and click the "Load" button.
  20. A "Completed !!" completion message is displayed.
  21. Click "OK".
  22. The vector sequence opened with the main feature map is loaded.
  23. Linearized DNA has end points for horizontal scrolling.
  24. Click "Cloning -> Ligation -> Simple Ligation ..." from the menu.
  25. The Ligation dialog will be displayed.
  26. There is a vector opened at the bottom of the list, and the shape of both ends is displayed.
  27. The end shape can also be seen in the sequence lane of the main feature map.
  28. Scroll horizontally until scrolling stops.

 

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