We did a mini prep experiment in lab where we isolated a plasmid and some of the

Question

We did a mini prep experiment in lab where we isolated a plasmid and some of the questions we didn’t discuss.
When the bacteria were mused under alkaline conditions, DNA and what were precipitated?
During the plasmid preparation, we used silica resin-containing microfilter column. What is the purpose of the silica?
Your DNA OD reading was 0.06 and dilution factor was 50. Determine the concentration of the undiluted sample?
Possible sources of error whilst executing plasmid purification. (The entire experiment was unsuccessful, technically, my professor and TAs were unsure of where the error occurred because everyone in class yielded negative results that were supposed to be positive. So I’m really not sure what to do here.) We did a mini prep experiment in lab where we isolated a plasmid and some of the questions we didn’t discuss.
When the bacteria were mused under alkaline conditions, DNA and what were precipitated?
During the plasmid preparation, we used silica resin-containing microfilter column. What is the purpose of the silica?
Your DNA OD reading was 0.06 and dilution factor was 50. Determine the concentration of the undiluted sample?
Possible sources of error whilst executing plasmid purification. (The entire experiment was unsuccessful, technically, my professor and TAs were unsure of where the error occurred because everyone in class yielded negative results that were supposed to be positive. So I’m really not sure what to do here.)
When the bacteria were mused under alkaline conditions, DNA and what were precipitated?
During the plasmid preparation, we used silica resin-containing microfilter column. What is the purpose of the silica?
Your DNA OD reading was 0.06 and dilution factor was 50. Determine the concentration of the undiluted sample?
Possible sources of error whilst executing plasmid purification. (The entire experiment was unsuccessful, technically, my professor and TAs were unsure of where the error occurred because everyone in class yielded negative results that were supposed to be positive. So I’m really not sure what to do here.)

Explanation / Answer

Ans.

Under alkaline conditions lipids are precipitated along with DNA.

The purpose of silica is that, DNA  binds to the silica membrane in the presence of chaotropic salts and under certain pH conditions. whereas, Polysaccharides and proteins do not bind well to the silica column. after binding and washing, nucleic acids are selectively eluted under low-salt conditions.

At O.D 1, the solution contains 50 micrograms of double stranded DNA per mL.

so at O.D 0.06 the solution will have 50*0.06 = 3 micrograms of double stranded DNA per mL.

as the solution is 50 times diluted, the concentration of undiluted sample is 3/50 = 0.06 micrograms of double stranded DNA per mL or 60 ng  double stranded DNA per mL.

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