So I am in a bit of a time constraint. Essentially, I inserted a DNA fragment via molecular cloning which contains a unique RE site. I need to confirm whether my colony has or does not have the fragment. I need to do this in one day so would this work as a confirmation strategy:
- Pick colony and perform colony PCR with primers around RE site.
- Cleanup the PCR product and digest using the enzyme which was inserted.
- Run on a gel. If a cut is noticed, this confirms that the colony had the inserted fragment (containing the RE site).
Would this strategy work? Are there any notable issues people can for see in this strategy?