Question

you are running a transformation reaction, and in your microfuge tube currently is your plasmid and your insert fragments (both containing compatible sticky ends). the other student worker who worked the previous lab shift, however, did not put the ligase back into the freezer, so it denatured and will no longer work, but you didn't know this at the time, and added it to your solution. explain exactly what will (or will not) happen now in your experiment, and how your transformation will be affected.

Answer 1

A foreign DNA molecule can be incorporated into a bacterial plasmid during a transformation reaction.

How to explain the reaction?

With the aid of two enzymes, ligase and restriction enzymes, a foreign DNA molecule can be incorporated into a bacterial plasmid during a transformation reaction. Each enzyme detects a target DNA sequence and cuts it nearby, while ligase aids in connecting the DNA. When two bits of DNA have complimentary bases, it facilitates their joining.

Plasmid and the insert fragment are both present in the microfuge tube, and they both have compatible sticky ends. However, the ligase has been denatured and is no longer active because the prior student left it outside rather than freezing it; despite this, we had already put the ligase into the tube. Ligase aids in binding the plasmid and insert fragments together, but because it is denatured in this instance, it will no longer be able to do so. As a result, no transformation process will take place. And since ligase links DNA fragments together by catalyzing the development of connections between the nearby nucleotides, the two fragments will not be able to unite.

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