Question

In this procedure, let us assume that the cell pellet volume is about 50 µL and you’ve added 100 µL of lysozyme solution, 200 µL of alkaline SDS and 150 µL of acidic sodium acetate (3M acetate, pH 4.8); therefore, the total volume is approximately 500 µL. Would it make any difference to the plasmid DNA yield if you altered these volumes? Explain your answer.

Answer 1

Yes, there will be a difference in the yield of DNA.

As lysosomes help in solublizing the peptidoglycan layer of the cell and sds, a detergent is used to solublize the proteins & lipids in cell membrane and also helps in making DNA histone free

If the concentration of these is less than the cell pellet then, all the plasmid DNA might not get free and thus, result in low DNA yield.