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- Question 1. Although we will not be doing a gel electrophoresis, data from a gel digest of a Bacillus anthrax plasmid is provided so you can do a DNA map. The Bacillus anthrax plasmid is 4000bp (4Kb) long. Note the origin position as well as the reference molecular weight markers on the gel. Two restriction enzymes, A and B, were used to obtain two individual digests, A and B. They were combined to produce the third digest. The restriction enzyme fragment pattern for the digest of Bacillus anthrax plasmid Determining the Number of Fragments How many fragments were produced by enzyme A? How many fragments were produced by enzyme B? How many fragments were produced by the combined digest (A and B)? Fragment Size Fragment size is relative to molecular weight, and must be determined by comparing the fragment distance to the molecular weight markers. The fragment size has been provided on the gel pattern for this exercise. To make a map you must determine the relative positions of the…QUESTION 5 Imagine that you have cloned the gene encoding the SARS-CoV-2 spike protein to a plasmid vector. The next thing is to put the plasmid into a host. Describe how you can transform the plasmid containing the gene encoding the SARS-CoV-2 spike protein.Question 1. Restriction endonucleases can be isolated from a number of bacteria. In bacteria restriction endonucleases. a-restrict chromosomal DNA that is heavily mutated b-restrict chromosomal DNA with significant regions that have been Deleted. c-restrict the DNA of invading bacteriophages. d- all of the above Question 2. In a PCR reaction the step in which DNA polymerase replicates the DNA is referred to as a- denaturation b- annealing C- extension d- initiation
- PLEASE CHECK THE WORDS THAT ARE IN BOLD. THEY ARE THE SAME QUESTIONS BUT THE WORDS IN BOLD ARE DIFFERENT. Assuming a linear piece of DNA (plasmid) was used as starting material, how many restriction sites were there considering you observed three bands after agarose gel electrophoresis. Assuming a circular piece of DNA (plasmid) was used as starting material, how many restriction sites were there considering you observed two bands after agarose gel electrophoresis.Question 3 PUC plasmids are widely used because of the beta-lactamase gene that renders an easier blue and white screening process in transformant selection. (A) True B) FalseQuestion 5: Try cutting Plasmid 1 with Xhol. Xbal. Kpnl, and Pstl. Try cutting with all 4 together." Would these enzymes be helpful and informative for creating a restriction map?-Why, or Why not?
- 5) Answers to the following questions. A. You are given a tube containing 275 ng of purified PCR product (DNA) that is 1262 bp long. How many picomoles of PCR product are in the tube? The average weight of a deoxynucleotide monophosphate is 328 g/mol. B. You digest a 5 kb plasmid with two enzymes that yield fragments of 4 kb and 1 kb, and resolve the digest on the gel. What would you predict to see in terms of the intensities of the bands?Question 7 Referring to the plasmid below, if a recombinant plasmid were obtained by inserting DNA into the EcoRv site, and the protein corresponding to the recombinant gene were expressed, which of the following statements would be false? Aval Sall A The plasmid will be resistant to tetracycline. B) The protein is not likely to be biologically active without some further treatment. C) The plasmid will be resistant to ampicillin. (D) The plasmid will not be able to replicate autonomously. Pul- Poul- ampr III EcoRI EcoRV BamHI pBR322 (4363 bases) -Poudl lettPlease answer both Part a. The plasmid you used as a template in your positive control for your PCR is 3300 bp in size. Why is the PCR product only 650 bp long? b. You using PCR to check for the presence of the GFP gene, however the thermocyder (machine that through temperatures) has been misprogrammed and tge annealing temperature is set to 43c (low for your primers) Explain what you would expect to see on your gel and why?
- The following image will be used to answer questions 1 through 9. Below is a restriction map of the circular plasmid YIP5. This plasmid contains 5,541 base pairs. There is an EcoRI site at base pair 1. The locations of other restriction sites are shown on the map. The numbers after the enzyme names tell at which base pair that enzyme cleaves the DNA. If you digest YIP5 with any one of the listed restriction enzymes you will end up with a linearized piece of DNA that is 5,541 base pairs long. EcoRI 1/5541 HindIII 32 Pvul 4916 Eagl 942 YIP 5 Apal 2035 Pvull 3247 Smal 2540 Reaction B) You digest YIP5 with two enzymes, Hindll and Apal, at the same time. How many DNA fragments would you expect from this reaction and what are the sizes of the fragments?The following image will be used to answer questions 1 through 9. Below is a restriction map of the circular plasmid YIP5. This plasmid contains 5,541 base pairs. There is an EcoRI site at base pair 1. The locations of other restriction sites are shown on the map. The numbers after the enzyme names tell at which base pair that enzyme cleaves the DNA. If you digest YIP5 with any one of the listed restriction enzymes you will end up with a linearized piece of DNA that is 5,541 base pairs long. EcoRI 1/5541 HindIII 32 Pvul 4916. Eagl 942 YIP 5 Apal 2035 Pvull 3247 Smal 2540 Reaction A) You digest YIP5 with two enzymes, EcoRI and Eagl, at the same time. How many DNA fragments would you expect from this reaction and what are the sizes of the fragments?Question 13 If a recombinant plasmid (below) was obtained inserting DNA into the BamHI site, screening for the recombinant plasmid can be done by which of the following technique? A) Plate on agar plates containing tetracycline and ampicillin. (B) Plate on agar plates containing ampicillin, C) Plate the cells on agar containing ampicillin then surviving colonies are plated on another agar plates with tetracycline. D) Plate on agar plates containing tetracycline. Pal Pord- ampr EcoRI ПРИ Ano pBR322 (4363 bases) Prull BamHI Sall let" Aval -Sall