ABSTRACT
In this session, we will run the large-scale digests from Experiment 4 on an agarose gel. Rather than just take a picture, however, this time we will physically remove the pieces and purify them from the agarose. We will then use these purified pieces in a ligation reaction to attempt to stick them together. e efficiency of DNA ligase is extremely small, so the resulting ligation product is not useful as is. Instead, we will transform the ligation reaction into highly competent Escherichia coli. If things go right, the bacteria will amplify only the desired ligation product and not any undesirable pieces, such as unrestricted vector, recircularized vector, and so on. e colonies will grow overnight and will be picked by students or instructors the following day.
