Question

why is it important that the DNA preparation be sterile /free of cells for Transformation experiment?

Answer 1

Ans. Transformation: A naked DNA is naturally taken up by the cell without mediation of any viral agent or artificial methods.

# The significance of using sterile environment for isolating cell-free DNA for transformation experiment are as follow-

I. Only the naked (cell-free) DNA is taken up by the host cell. Therefore, the required DNA (gene of interest) must be present in a cell-free medium.

II. Presence of contaminants (like, sweat, bacteria, dust, etc.) may lead to degradation of desired DNA by the nucleases present in the contaminants. So, all the DNA-preparation protocol must be carried out under sterile conditions.

III. Presence of contaminants may also adversely affect the host cells to be transformed (for example, the host cell can be infected by virus or bacteria, etc.) and reduce the overall transformation efficiency simultaneously with causing unexpected/undesirable outcomes in the experiment.