Talk:BE.109:DNA engineering/DNA ligation and bacterial transformation
You can use the plasmid maps that are provided with the protocols to choose some suitable enzymes for your diagnostic digests. There is no need to use the web-based NEB cutter tool, which has been unavailable periodically tonight (02/27/06).In fact, if you go with the plasmid maps that are included with the class material, you'll be more likely to come up with enzymes the lab actually owns since the more common enzymes are the ones that are listed. Post comments, problems or suggestions here. Good luck -- Natalie
To help you through the logic in planning your ligation, please take a look at the attached slide. Note that for your homework the length ratio is described as 1:4 - this is my mistake, it is actually ~1:7. If you have already done your calculations based on the 1:4 ratio, it is fine to go ahead with what you have planned.
Hopefully the slide below will help you think through the logic of how to set up a reaction with a reasonable ratio of backbone:insert (a ratio of 1:3.5 is fine!).
--Prof. Engelward
