Exhausted?

A good night's rest will do wonders.

We hope you will still remember some of this stuff tomorrow!
To make sure, we'll summarize briefly what we did:

We wanted to transfer GOI from pGEM-GOI to pQE60.

At Home

We obtained the sequence of pGEM from the company selling it. We then drew the plasmid pGEM-GOI and studied its orientation. Then we analysed its restriction map and the ATG location.
The PDF file describing the plasmid contains important information. Always check it!!! Sometimes there is a mistake in the numbering of the ATG or the MCS position. Don't trust anybody! Another thing: The restriction sites given in the PDF file are only part of the whole restriction map. (Usually they give the most frequently used). Always check for additinal sites using WebCutter or another software.
We then obtained the same data for pQE60.
In Class
We fed this data to the CloneIt software, and received suggestions of various strategies to perform this cloning. The computer helps a lot, but it does not think instead of you!!! First make sure you enter the correct data!!! One slip can give a great strategy for something you don't want or can lead to the impression that such a cloning is impossible. Always check the solution.Then use your common sense to evaluate the differnt strategies and choose the one best for your purpose.

We hope you learned some new things that may help you in future.

Your comments are most welcome.