We have three plasmids: the cloning vector, pGEM, the one that result if BamHI h

Question

We have three plasmids: the cloning vector, pGEM, the one that result if BamHI had been used to construct the genomic library, and the one that would result if EcoRI had been used to construct the library. What is the expected number of DNA fragments that would result from digesting...

pGEM with the restriction enzymes BamHI, EcoRI, and BamHI + EcoRI?

pGEM + BamHI insert with the restriction enzymesBamHI, EcoRI, and BamHI + EcoRI?

pGEM + EcoRI insert with the restriction enzymesBamHI, EcoRI, and BamHI + EcoRI?

Explanation / Answer

In case 1. pGEM with the restriction enzymes BamHI, EcoRI, and BamHI + EcoRI pGEM (cloning vector) digested with BamHI - will give only one fragment because BamHI is cut only at one site so it will linearized the circular plasmid Similarly, pGEM digested with EcoRI -will give only one fragment. pGEM digested with BamHI + EcoRI - will give two fragments because both bamHI and EcoRI will cut and produce two fragments out of circular plasmid Case2. pGEM + BamHI insert with the restriction enzymesBamHI, EcoRI, and BamHI + EcoRI pGEM + BamHI insert with the restriction enzymes BamHI - will give two fragments pGEM + BamHI insert with the restriction enzymes EcoRI will give single fragment pGEM + BamHI insert with the restriction enzymes BamHI+ EcoRI will give three fragments Case3. pGEM + EcoRI insert with the restriction enzymesBamHI, EcoRI, and BamHI + EcoRI pGEM + EcoRI insert with the restriction enzymes BamHI- will give only one fragment pGEM + EcoRI insert with the restriction enzymes EcoRI - will give two fragments pGEM + EcoRI insert with the restriction enzymes BamHI+ EcoRI- will give three fragments because BamHI cut at single site and EcoRI cuts at two site (both at end of insert)

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