The rate of migration of DNA within an agarose gel in the gel electrophoresis technique is dependent on what factor(s)? Select one or more: a. G-C / A-T ratio b. Well size of the gel c. Size of DNA fragment d. Volume of sample loaded e. Negative charge of DNA
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- The rate of migration of DNA within an agarose gel in the gel electrophoresis technique is primarily based on: O The number of the DNA fragments. The size of the DNA fragments. The concentration of the DNA NoneWhy is the company Qiagen has more refined DNA extraction steps than a normal Strawberry DNA extraction practical? Summary of Qiagen DNA extraction steps Add ATL buffer and grind with sample. Add 20 microliters of enzyme Proteinase K to degrade protein into a 1.5-2ml microcentrifuge tube. Add 200 microlitres AL lysis buffer, and mix by vortexing for 5–10 seconds, which breaks cell membrane allowing DNA to be released. Incubate the sample at 56 degrees for 10 minutes. Mix the cell lysate with 200 microlitres ethanol by pipetting it at the side of the microcentrifuge wall so DNA precipitates. The DNA forms a white layer and the remaining liquid is discarded. Pipet the mixture into DNeasy Mini spin column placed in a 2 ml collection tube. Centrifuge for a minute at 8000 rpm. Place the mini spin column into a 2 ml collection tube, add 500 µl Buffer AW1, and centrifuge for 1 min at 8000 rpm. Then add it to a new 2 ml collection tube (provided), add 500 µl Buffer AW1, and centrifuge for 1…You digest 4 uL of plasmid DNA that is 50 ng/uL concetration in a total volume of 20 uL. You run 10 uL of the digest on teh gel. You then do a DNA purification protocol with a Zippy prep on the remaining digested DNA. You elute the DNA in a 25 uL. A 2 uL ssample has the concentration of 2 ng/uL. What is the DNA yield? YIELD = How much dna came out of the column/how much DNA was loaded onto the columnRemember to subtract amount of DNA run on gel from total digested DNA to get hw much DNA was loaded onto the columnAmount that came out of the column equals the volume of the eluted DNA times the concentration of the purified DNA
- 4. Look at the gel image and answer the questions below and be specific. a) Based on your calculations of the DNA concentrations, how much DNA was loaded into each well? Do you see DNA for each of your samples? If not, why do you think that is so? b) Is the DNA in a single sharp band, multiple bands or a smear? What would each of these scenarios be due to, and why would you see them for your samples? c) Do you see multiple bands in your plasmid DNA sample? What are they?Which of the following best describes the process of DNA seqencing. a. DNA is seperated on a gel and the different bands are labled with flouroscent nucleotides and scanned with a laser. b. A laser is used to flurorescently label the nucleotides present with in the DNA , the DNA is run on a gel and then the DNA is droken into fragments c. Nucleotides are scanned with a laser and incrprorated into the DNA that has been seperated on a gel and then DNA is amplified with PCR. d. fragments of DNA are produced in a reaction that lables them with any of four different fluroscent dyes and the fragmented then are run on a gel and scanned with laser e. DNA is broken down into its constituents nucleotides and the nucleotides are then run on a gel and purified with a laserWhich of the following statements is TRUE about buffers in the separation of DNA strands via gel electrophoresis? O The running buffer contains ions that allow electric current which moves the DNA bands in the gel O The loading buffer helps in the cleaner separation of DNA bands during electrophoresis O Tris-borate-EDTA has a better buffering capacity than Tris-acetate-EDTA (TAE), that is why it is used in the separation of larger DNA strands O TAE is more heat resistant than TBE.
- During gel electrophoresis, in what direction (toward which electrode) does DNA move toward? O positive O negative O both O either onePlease answer the following using the experiment data below. Provide the formulas or methods used for calculations of each. Number of colonies on LB/amp/ara plate = Micrograms of DNA spread on the plates = Transformation efficiency = DNA plasmid concentration: 0.08 μg/μl 250 μl CaCl transformation solution 10 μl pGLO plasmid solution 250 μl LB broth 100 μl cells spread on agar 227 colonies of transformantsExamine the pGLO plasmid DNA solution with the UV lamp. Note your observations. Using a micropipettor, withdraw 10 ul of plasmid and mix it into the cell suspension of the +PGLO tube by pipetting up and down three times. Close the tube and return it to the rack on ice. Also close the -PGLO tube. Do not add plasmid DNA to the -pGLO tube. Why not?
- In your forensic laboratory, you set up a reaction to digest DNA with restriction enzymes. The final volume will be 20 ?μl. How much of each of the following reagents will you use to obtain the indicated working concentrations? 10x restriction buffer >> working concentration 1x 1 mg/mL bovine serum albumin (BSA) >> working concentration 100?μg/mLThe 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.When gel electrophoresis is undertaken, one well always contains DNA-marker molecules. The purpose of these markers is to Question 30 options: help visualize the DNA indicate when the DC current is turned on serve as a guide to the length of the fragments in the other wells indicate when the DNA has completely crossed the gel serve as a channel for ethidium bromide