Virtual Cloning Tutorial - Human Proinsulin

Open the Human Proinsulin sequence.

Using Find Restriction Sites you're going to locate the NcoI site. Make sure that the Must not cut between bases box is unchecked this time. Still using the commonly used enzymes, click the Advanced button to bring up the list of enzymes. Scroll down to find NcoI and select just that one then click Done.


This will annotate just one NcoI site near the beginning of the CDS. Click Apply and save the document.

Next we are going to introduce a cut site at the 3' end of the gene using PCR.


Exercise 4: Cloning Primer Design
Exercise 5: Introduce Cut Site using PCR
Exercise 6: Insert into Vector