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Electrophoresis is an extremely useful procedure when applied to analysis of
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Concepts of Genetics (12th Edition)
- How many microliters of the pGLO plasmid do you need if you want 5 micrograms of DNA and the plasmid concentration is 100 nanograms/microliter?arrow_forwardTotal nucleic acids are extracted from a growing culture of yeast cells. They are then mixed with specialized beads to which the single-stranded DNA molecule with sequence 5’-TTTTTTTTTTTTTTTTTTTTTTTTT-3’ has been covalently attached to the surface (see image to the right, where each black line represents a polynucleotide sequence). After a short incubation time, the beads are removed from the mixture. When you analyze the cellular nucleic acids stuck to the beads, which type of nucleic acid (i.e. DNA, rRNA, etc.) do you expect to be the most abundant? Why?arrow_forwardCoomassie dye is the equivalent of what component used in analysis of DNA by agarose gel electrophoresis? How do you expect a Coomassie-stained gel to look compared to the result you might expect once you have completed your Western blot?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_forwardYou have isolated genomic DNA and plasmid DNA from a bacterial culture. Please answer the following questions: 1. What solution did you use to elute DNA from the silica resin column? 2. What is the function of the chaotropic salt in DNA isolation? 3. Why do we add the chaotropic salt for genomic DNA before lysis and for plasmid isolation we add the salts after the lysis step?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
- In DNA purification, explain how the chromosomal DNA is separated from the plasmid DNA? Be sure to mention the specific buffer components that facilitate this process.arrow_forwardIs a faded gel matrix in a DNA ladder in agarose gel electrophoresis still useable for analysis or not?arrow_forwardIT IS POSSIBLE TO ESTIMATE THE CONCENTRATION OF SPECIFIC DNA SEQUENCE IN A SOLUTION OF MIXED DNA. Explainarrow_forward
- The following image is of an agarose gel. If DNA samples were loaded to this gel and the electrophoresis experiment was started, explain what would happen and why.arrow_forwardYou add 3.0 µg of plasmid DNA to a QIAprep spin miniprep purification column. You perform the binding in a binding buffer at pH 6.8. What portion (%) of the 3.0 µg of plasmid DNA would you expect to bind to the column in these conditions? What portion (%) of the bound plasmid DNA would you expect to elute using elution buffer at pH 8.5? Make the following assumptions:• Solid phase silanol groups can have a very wide range of pKas. For the purpose of this problem, assume the silica gel functional group (silanol) in the DNA purification column matrix has a pKa of 7.5.• If a DNA molecule and column matrix functional groups have the same charge state, then that DNA molecule will be repelled and will not bind to the column. If they do not have the same charge, binding occurs.arrow_forwardWhich of the DNAs shown in Figure would move fastest during agarose gel electrophoresis?arrow_forward
- Human Heredity: Principles and Issues (MindTap Co...BiologyISBN:9781305251052Author:Michael CummingsPublisher:Cengage Learning