Question

When referring to MO BIO Microbial DNA Isolation kit protocol...

8. a) How is the DNA separated from the rest of the cell debris after the lysis step?

b) Where will the micro-beads be once the DNA is separated from the cell debris?

Answer 1

a) How is the DNA separated from the rest of the cell debris after the lysis step?

Answer: Isolation of DNA with MO BIO Microbial DNA Isolation kit protocol:

Figure: Flow chart showing microbial DNA isolation procedure using MO BIO Microbial DNA Isolation kit.

The Solution MD1 contains SDS (Sodium dodecyl sulfate or sodium lauryl sulfate) and other disruption agents required for cell lysis. In addition to helping in cell lysis, The SDS is an anionic detergent that breaks down fatty acids and lipids associated with the cell membrane of several organisms.

The Vortex creates the combined chemical/ mechanical lysis conditions required to release desired nucleic acids from microbial cells. The chemically enhanced bead beating process helps in cell lysis. Without this chemically enhanced beading beating process many cell types will not lyse. The vortex action is typically all that is required for cell lysis along with micro-beads.

After lysis the sloution DNA is present in the supernatant while the cell debris gets precipitated at the bottom of the micro-bead tube. The supernatant is then transfered to collection tube. The supernatant to be collected very carefully avoiding any pellet or cell deris.

b) Where will the micro-beads be once the DNA is separated from the cell debris?

Answer: After cell lysis the DNA gets seperated from the cell debris. The DNA is present in the supernatant. while the cell debis and micro-beads remain in the bottom of the microbead tube.