You purified the plasmid from an overnight cell culture and dissolved the plasmid DNA in 100...

You purified the plasmid from an overnight cell culture and dissolved the plasmid DNA in 100 μL 0.1M Tris buffer (pH8). To quantify the plasmid, you took an absorbance measurement at 260 nm of a 1:50 dilution sample. The absorbance was 0.35. What is the concentration of the undiluted plasmid in micrograms per mL?

Expert Solution

Ans. The average extinction coefficient of a double stranded DNA is 0.020 ug mL^-1cm^-1 that gives the value of 50.0 ug/ mL DNA concentration for absorbance of 1.000 at 260 nm.

So, an absorbance of 1.000 at 260 nm indicates 50 ug/mL DNA- it is taken as the standard or reference.

Concentration of DNA solution with absorbance of 0.350 at 260 nm=

(Abs. of test solution / Abs. of standard) x concentration of standard.

= (0.350 / 1.000) x 50.0 ug/ mL

= 17.5 ug/ mL

Thus, the concentration of 1:50 diluted sample = 17.5 ug/ mL

Now,

Concentration of original undiluted solution = concentration of diluted sample / dilution factor

= (17.5 ug/ mL) x (1 : 50) = (17.5 ug/ mL) x (1 / 50)

= 875 ug/ mL

Thus, concentration of undiluted plasmid = 875 ug/ mL

queen_honey_blossom answered 1 year ago

A virus particle is part of a cell True or False? Choose which purified DNA will...

A virus particle is part of a cell True or False? Choose which purified DNA will lead to a productive infection in a transfection experiment. A) single stranded DNA from herpes virus type 2 B) single stranded DNA from poxvirus C) single stranded DNA from picornavirus D) single stranded DNA from parvovirus. E) single stranded DNA from papovavirus. A virion is A) an infectious protein B) an infectious virus particle C) a virus D) an infectious nucleic acid E) an...

Assume you have purified the insert-containing plasmid from chloramphenicol or kanamycin-resistant E. coli. You used pBLU...

Assume you have purified the insert-containing plasmid from chloramphenicol or kanamycin-resistant E. coli. You used pBLU as the vector for the unknown insert project. You use PCR to identify your unknown insert. What is your positive control, negative control, and experimental group?

2) Please explain how to purify plasmid DNA from bacterial cells.

You derive a cell line from a tumour and discover that it is deficient in DNA...

You derive a cell line from a tumour and discover that it is deficient in DNA helicase activity and DNA repair activity. Is this helicase deficiency likely to affect base excision or nucleotide excision repair? Explain your reasoning.

1) A plasmid has the antibiotic resistance genes for kanamycin and ampicillin. You insert foreign DNA...

1) A plasmid has the antibiotic resistance genes for kanamycin and ampicillin. You insert foreign DNA into the ampicillin resistance gene. E. coli containing the recombinant plasmid (plasmid + foreign DNA) will grow in the presence of ampicillin but not in the presence of kanamycin. A)True B)False 2)Expression of a gene could result in... A) transcription of a mRNA from the gene B) transcription of a protein from the gene C) translation of the DNA of the gene 3)Restriction enzymes...

If you estimate that an overnight culture has 5.28x10^7 viable colonies per mL, how would you...

If you estimate that an overnight culture has 5.28x10^7 viable colonies per mL, how would you confirm this? Please draw ALL dilutions and write what dilution each tube and plate represents and how many bacteria you expect per mL in each tube. Write down how many bacteria will be on the final plate. Making sure that the number falls within the expected range for accurate counts.

I want you to research how to purify a protein from cell culture. After purification of...

I want you to research how to purify a protein from cell culture. After purification of the protein, you would need to quantify the protein using Bradford Assay and run the protein on SDS-PAGE gel for Western blot analysis to determine the amount of specific in a sample. Write out a protocol for the isolation of protein from human cell culture- you can determine the type of cells you are purifying the protein from Write out a protocol for the...

Describe how bacterial DNA transfer from one cell to the others.

You are monitoring growth of a culture by taking 100 microliter samples from 1:1000 dilutions of...

You are monitoring growth of a culture by taking 100 microliter samples from 1:1000 dilutions of the culture. At T0 plate count was 80 colonies, at T1(20) it was 120, at T2(40) is was 160. Give the CFU/ml of the culture for each time and indicate what was the doubling rate per hour of the organism.

You have an E. coli culture of which you are trying to determine the cell concentration....

You have an E. coli culture of which you are trying to determine the cell concentration. You need to perform a serial dilution. You take 100 µL of the undiluted stock culture, add it to 9.9 mL water in tube A, and mix. Then you take 100 µL of the material in tube A, add it to 9.9 mL water in tube B, and mix. Then you take 100 µL of the material in tube B, add it to 9.9...

Question