Give a short discussion for each of the reagents of the plas

Give a short discussion for each of the reagents of the plasmid isolation procedure. Include the step the reagent is associated, and the function of the reagent (in relation to the DNA/proteins/ or other chemicals)* Note- not all the reagents are provided for each buffer/solution used in the plasmid Isolation experiment. Include the resource(s) that you use to help answer this question.

70% Ethanol -

200mM sodium Hydroxide/1% SDS -

Solution

Cultivation of the bacterial sample that contains your plasmid DNA. An antibiotic is usual incorporated in the growth media and the plasmid DNA will pass resistance to the antibiotic on to the bacteria.

Resuspend the pelleted cells in buffer solution - solution containing Tris and EDTA chelates divalent metal ions such as magnesium and calcium. This destabilizes the cell walls and inhibits the action of DNases. Adding glucose to the buffer solution helps maintain osmolarity to keep the cells from bursting while adding.

A highly alkaline solution consisting of NaOH and SDS is then used to break down the cell walls and convert the double stranded DNAs (dsDNA) to single stranded DNAs (ssDNA).

Neutralize the solution with potassium acetate: adding KAc to the solution, you make it possible for the ssDNAs to re-nature into dsDNAs. These double stranded plasmid DNAs are easily dissolved in the solution while the genomic DNAs (gDNA), denatured cellular protein and SDS will stick together to form a white precipitate that can easily be separated through centrifugation.

Isolate your plasmid DNA by using a process known as ethanol precipitation. Rinse the resulting precipitate (your plasmid DNA) in ice-cold 70% ethanol and let it dry for about 10 minutes to allow the alcohol to evaporate.

You should likewise resuspend the DNA pellet in a buffer solution containing Tris, EDTA and RNases to cleave all the remaining RNAs in the solution