Question

You've performed a shotgun cloning experiment with your favorite organism and pGEM 7 as the cloning vecto.

After the ligation and transformation, you observe a plate with a mixture of blue and white colonies

If you isolated plasmid DNA from blue colonies what would you expect to observe in relation to the presence of an insert? And if you isolated plasmid from white colonies what would you expect to observe in relation to the presence of an insert?

Answer 1

As we know that cloning means to get multiple same copies of particular DNA or anything which we want to clone.

Shot Gun cloning is a type of cloning method which is very slow having maintenance issue and also clone bias so it not used rapidly.

In shot gun cloning method

1. Large DNA molecules cut into small fragments

2.fŕagments inserted into plasmid vector like that here is pGEM7

3.After that Sequencing Genome all done and we run it on computer to get result.

After all that process as we got blue and white colonies there are several reasons to get blue and white colonies are as follows:

First is if plasmid is non recombinant will turn into blue colonies and recombinant plasmid will turn into white colonies.

Another one is if a plasmid containing functional Galacosidase gene inside him will turn in blue and this activity called alpha implementation

And if Beta Galacosidase gene disrupt in plasmid than it will turn into white colonies.