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Electrophoresis is an extremely useful procedure when applied to analysis of
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Concepts of Genetics (12th Edition)
- A piece of DNA is cut into four fragments as shown below. A solution containing the four fragments is placed in a single well at the top of an agarose gel. Using the information given below, draw (below the well) how you think the fragments will be aligned on the gel following electrophoresis. Label each fragment with its corresponding letter. Remember, each band on the gel will be the same width, equal to the width of the well at the top of the gel. These should all be in one lane. What if you had two different DNA fragments that were exactly the same length as measured in base-pairs. Would it be possible to distinguish them using this type of electrophoresis? How would they appear on a gel?arrow_forwardPlease briefly explain what gel electrophoresis is and how it works to separate a mixed sample of macromolecules like DNA.arrow_forwardBoth protein and DNA are run together in an isoelectric focusing (IEF) electrophoresis using the immobilised pH gradient (IPG) strip with pH range of 4-7. After the electrophoresis and staining, only ONE band is observed on the middle of the IPG strip. The band is a protein band. Briefly explain why only the protein band and NOT the DNA band appear on the IPG strip.arrow_forward
- At what stage of the culture should bacterial colonies be harvested for plasmid DNA extraction? How about for genomic DNA extraction? What distinguishes the xanthogenate-based methodology from the traditional phenol/chloroform method for isolating DNA from bacteria? What makes potassium ethyl xanthogenate efficient in isolating DNA from a variety of microorganisms?arrow_forwardDiscuss the use of gel electrophoresis for the separation of macromolecules (DNA, RNA and protein) of different sizes and topological forms. (Include in you answer the different recombinant DNA techniques that require gel electrophoresis)arrow_forwardYou are about to isolate a 3000 bp large plasmid from an E.coli culture. You know that the plasmid is present in 100 copies per E. coli cell. You aim to have a final plasmid concentration of 100 ng/µl in a total volume of 50 µl. Assuming the yield is 100 %, how many E. coli cells should the culture from which the plasmid is to be isolated at least containarrow_forward
- Discuss the use of gel electrophoresis for the separation of macromolecules (DNA, RNA and protein) of different sizes and topological forms. (Include the different recombinant DNA techniques that require gel electrophoresis)arrow_forwardYou are about to isolate a 3000 bp large plasmid from an E.coli culture. You know that the plasmid is present in 100 copies per E. coli cell. You aim to have a final plasmid concentration of 100 ng/µl in a total volume of 50 ul. Assuming the yield is 100 %, how many E. coli cells should the culture from which the plasmid is to be isolated at least contain A) 4.7 · 1013 cells B) 1.6 · 1010 cells C) 3.2 · 1010 cells D) 3.2 · 1026 cells E) 1.6 · 1012 cells Solution B that was used during the plasmid isolation contains 0.2 M NaOH (see practical manual). Which effect does NaOH have on E. coli DNA A) It denatures genomic DNA and plasmid DNA. B) It denatures genomic DNA and but not plasmid DNA. C) It denatures plasmid DNA but not genomic DNA. D) It denatures neither genomic DNA nor plasmid DNA. E) This is unpredictable. Which statement on the migration of DNA fragments through agarose gels is false A) Small fragments migrate faster than larger fragments because they can move faster through…arrow_forwardUsing a ThermoFisher GeneJet Miniprep plasmid isolation kit to isolate plasmids from bacteria: Use of RNAse increases purity of purified plasmids. This enzyme will be added when the cells are lysed, detroying all RNA in the sample. Why will this action increase the purity of an isolated plasmid sample? This will only be used for the plasmids isolated using the GeneJet purification columns, and not when performing alkaline lysis. How will this affect the predicted results for the alkaline lysis plasmid isolations?arrow_forward
- You have purified plasmid DNA, and observed the following absorbances: A260 = 2.9 A280 = 2.2 If A260 of 1.0 = 50 µg/ml pure dsDNA. Which of the following can you conclude? Select all that apply The concentration of your plasmid DNA is 145 ng/ul. The concentration of your plasmid DNA is 110 ng/ul. The plasmid DNA purity is acceptable. The plasmid DNA purity is not acceptable.arrow_forwardWhat is the structure of the gel stain being used? There are several other dyes that can bind DNA and used for gel imaging. Find an example of such a dye, and draw its structure.arrow_forwardYou're purifying some plasmid DNA from a culture of bacteria and you want to know how pure it is. You measure the optical density at 260 m and 280 m and find the ratio is 2.0. You suspect there is RNA contamination in your preparation, so you treat your preparation with RNase. But the ratio is still 2.0. Protein assays tell you there is no protein in your solution, and no other biological molecules absorb light very efficiently at those wavelengths. What's the explanation?arrow_forward
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