Give typing answer with explanation and conclusion The use of spin columns with silica for plasmid purification comprises an example of the following purification technique: a. adsorption to a matrix b. non-organic extraction c. organic extraction d. alkaline extraction e. none of the above
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Give typing answer with explanation and conclusion
The use of spin columns with silica for plasmid purification comprises an example of the following purification technique: a. adsorption to a matrix b. non-organic extraction c. organic extraction d. alkaline extraction e. none of the above
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- 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.Please 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 transformantsChoose the proper order for recombinant protein purification from bacterial cells. a. lysis,pelleting by centrifugation, induction of protein expression, transformation, purification by chromatography b. induction of protein expression, transformation, pelleting by centrifugation, lysis, purification by chromatography c. transformation, induction of protein expression, pelleting by centrifugation, lysis, purification by chromatography d. purification by chromatography, lysis, pelleting by centrifugation, induction of protein expression, transformation
- When plasmids are isolated from bacterial cells, they may existin a number of forms.a. List the different forms that may be found.b. Which do you think would migrate the fastest and farthest in anelectrophoresis experiment and why?Please briefly explain what gel electrophoresis is and how it works to separate a mixed sample of macromolecules like DNA.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.
- 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.What should be the first step in the protocol for the purification of DNA from any source (including human cells)? a. Phenol extraction b. SDS precipitation c. Ethanol precipitation d. Digestion of protein with Proteinase K e. RNase digestiondescribe the two main parts of a plasmid DNA extraction and purification procedure including the purpose of each step
- Use the plasmid map for pBashi below to answer the following questions Xhol Hind!!! 15 kb 11 kb 10 kb 8 kb 7 kb 5 kb a. How large is pBashi? b. On the gel below, draw the bands corresponding to the expected pBashi cleavage products after restriction digest with the indicated restriction enzymes (Lane 2 - Xhol only, Lane 3 - HindIII only, Lane 4 - Xhol+ HindIII) 4 kb 3 kb 3 kb 2 kb 1 kb 4 kb Hindill 1 kb 7 kb Xhol Xhol Xhol Xhol Hind!!! Ladder Xhol Hindill Hind!!! Pstl Pstl c. Lane 5 in the gel above displays the pBashi cleavage products after restriction digest with the restriction enzymes Xhol+ Pstl. Lane 6 in the gel above displays the pBashi cleavage products after restriction digest with the restriction enzymes Xhol+ HindIII + Pstl. Indicate your final answers to the questions below clearly on the pBashi plasmid above; you will NOT receive any credit if you only list sizes of fragments below. If you need to, draft your answer somewhere else before writing your final answer above.…You are provided with cultures of E. coli cells that contain the plasmid pUC118. Each student should carry out the plasmid isolation procedure. Note the appearance of the suspension after steps (v), (vi) and (vii). The steps are (v). Resuspend the bacterial pellet in 300 μl solution A [25 mM Tris-HCl (pH 8.0) 10 mM EDTA. Ensure that the suspension is homogeneous. (vi). Add 300 μl solution B [1% (w/v) SDS 0.2 M NaOH] and mix thoroughly. (vii). Add 300 μl solution C [3M potassium acetate, (pH 4. 3)], mix thoroughly, and incubate on ice for 5 minutes.The principle of the alkaline method for plasmid DNA isolation is: O A. An acidic solution differentially denatures genomic DNA and plasmid DNA B. Differential re annealing capacity of plasmid vs genomic DNA after denaturation with a basic solution O C. All the options are correct O D. Genomic and plasmid DNA are differentially washed by 70% ethanol O E. The action of the detergents on the cell wall