Explain the step in detail of method if want to insert targe

Explain the step in detail of method, if want to insert target geneaprX into vector. Then, the recombinant vector will be transferred to E. coli and how to select the successful ligation

Xba Kpn EcoR Sac BamH EcoR Inserted DNA Pst Pst Sac aprY

Solution

I will choose to digest pUC19 with EcoRI, BamHI, and SalI as these are the cheapest, good and commonest enzymes with unique sites good for E. coli. I\'ll digest the vector then incert my gene of interest by ligating with T4 DNA ligase and ATP for overnight. This will make a directional cloning of my gene of interest that will be in right orientation under promoter. I will transform competent E. coli cells like DH5alfa then clone the desired recombinant plasmid vector and will use further.