The student finally sets up the PCR, runs it on the gel and was able to determin

Question

The student finally sets up the PCR, runs it on the gel and was able to determine that they successfully amplified the Evan1 gene, which was about 1kb long. It tuns out that the gene was flanked on either side by a site recognized by the EcoRI restriction site. The student uses the EcoRI enzyme to cut the ends of the gene at those site and then proceeds to clone it into the pre-cut plasmid. The student mixes the cut gene fragment and the plasmid and remembers that they need one imprtant component to ensure that the ends stick together. What is that component?

Explanation / Answer

The component is T4 DNA Ligase. It is a very important component for DNA ligation reaction. It works in the presence of T4 DNA Ligase Buffer, and it is responsible to ligate the insert with the pre-cut plasmid.

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