CRISPR answer sheet experiment 1

a , b , c question

Part 1. Gene Cloning   1) How is a gene inserted into a plasmid cloning vector? Be specific, but be brief. No more than 2 (non-run-on) sentences please, and do not exceed this space.             2) A geneticist has transformed a vector with an ampicillin resistance gene into E. coli cells and plated those cells on media containing ampicillin.  She comes back 24 hours later and sees only blue colonies on the media. a) How does she know that the cells growing on the media were successfully transformed with the vector?           b) What does it mean that the colonies are blue?

A amp PBR322 4301 fot B Clear Zones Figure 2 The postgraduate student, Demika, inserted her gene of interest into the plasmid, pBR322, before transformation into the competent host cell using heat shock method. After that she cultured the cells on the Ampicillin agar plate before replica plating the colonies onto another Ampicillin (A) and Tetracycline (B) agar plates shown in Figure 2. (1) Referring to the vector pBR322 in Figure 2, which recognition site was cleaved to insert the gene of interest? Based on the observation above, can you identify which colonies are carrying positive mcombinants of BR322? Explain your selection.

3A.  Following transformation with the CRISPR plasmid, the yeast will be plated on SD-URA plates.  Why is SD-URA media used? Name the relevant marker gene in the pCRCT CRISPR plasmid.       3B. What would you expect to see if you plated on YED accidentally? Will most of the yeast be red or white? Why?       3C.  The complete genotype of the host yeast strain is MATa ade2 his3 leu2 met15 ura3.  Name one advantage of using a yeast strain with auxotrophies for several genes (i.e. his3, leu2 and ura3).         3D. After successful CRIPSR editing, the yeast can later be “cured” of the recombinant CRISPR plasmid – that is, the plasmid is lost but the CRISPR edit is stably inherited.  Write the new genotype of the yeast-based on the ADE6 gene.

Part ii. ) please

4. Animation 12.4: This table shows results from a genetic test conducted on DNA samples from two patients. The patients are a newly married couple seeking genetic counseling about their risks of passing on a genetic disease to any children they may have in the future. The samples included the DNA region including and immediately surrounding a single gene. Restriction enzyme cleavage analyses were performed on the samples. Female Patient 1) 3400 bp fragment Male Patient 1) 1600 bp fragment 2) 1800 bp fragment 3) 3400 bp fragment DNA fragments on gel What do the genetic test results indicate about this case? O Both the woman and the man are homozygous for the mutation. O Both the woman and the man are heterozygous. O The woman is homozygous for the mutation and the man is homozygous for the normal gene. O The woman is homozygous for the mutation and the man is heterozygous.

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This is question is from my micro biology class. 1. OUTLINE the steps that lead to the production of proteins from the genome of a double stranded RNA virus. List any PROCESSES involved and any ENZYMES.

1) Look at the ideal results. Were your predictions accurate, and how did they compare with your results?   2) You used aseptic technique during this lab. Why is it important to work in a sterile manner when working with bacteria in the lab?   3) Why are the cells incubated at 42°C?

7) Why are the following reagents used? Neutralizing solution (Plasmid isolation)    Isopropanol (Plasmid isolation)     RNase (isolation of genomic DNA)

Activity 1.3: Complete the table.Directions: Explain and give application for each process/tools.

1. Please help answer this question and please show all work on how you got the answers. Thank you so much!! :)

1) Given the image below, describe what is happening at each of the 3 sections (i.e. 1, 2, and 3) . [In section 1, the gene donor is providing..., In section 2..., In section 3...]

1a.The complete genotype of the host yeast strain is MATa ade2 his3 leu2 met15 ura3.  Name one advantage of using a yeast strain with auxotrophies for several genes (i.e. his3, leu2 and ura3). b.)After successful CRIPSR editing, the yeast can later be “cured” of the recombinant CRISPR plasmid – that is, the plasmid is lost but the CRISPR edit is stably inherited.  Write the new genotype of the yeast based on ADE6 gene.

1 2. 3. Two different cultures of Escherichia coli bacteria were inoculated onto nutrient agar plates. One culture was transformed with pGLO plasmid and the other culture was not. All of the plates shown above are illuminated with ultraviolet light. • Which plate was inoculated with E.coli+PGLO bacteria and included ampicillin antibiotic but did not include arabinose? • Which plate was inoculated with E.coli+pGLO bacteria and included both ampicillin antibiotic and arabinose. • What would you expect to observe on a plate that included ampicillin and was inoculated with E. coli minus (without) PGLO plasmid?

6. Explain why a positive test for COVID19 would appear sooner than a negative result when using real- time PCR to test if someone is infected. 7. Explain why the same primers (GMM) could detect several types of genetic modifications to common crop plants, rather than one specific gene or genetic modification. 8. Explain the general procedure for using the NanoDrop machine in analyzing DNA samples. Be sure to explain the purposes of the blanks (pure water), and of the importance of the 260/280 ratio. Identify an ideal 260/280 ratio of purified DNA.

10. When looking at your agar plate, where are the bacteria that did not take up the plasmid during the transformation? (It is a 4 letter word). Thanks for the help!

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Row C D. B B. one: A C O phage with radiolabelled protein coat phage with radiolabelled DNA 100 10. phage infects The experiment shown above was designed by bacterium phage infects bacterium EXPERIMENT 1 phage shell is removed EXPERIMENT 2 요 Hershey & Hershey & Chase Meselson & Stahl Meselson & Stahl 8 phage shell is removed 28 no radioactivity in cells ii 48 radioactivity in cells LL and proved that ii DNA replication is semiconservative DNA is the hereditary material DNA replication is semiconservative DNA is the hereditary material (select the row that correctly completes the statement)

A cloning vector map is shown below. EcoRI Bam Ban Hind P-galactosidase Amp Bam Bam EcoRI Ori C Which restriction site is best for inserting a DNA fragment for selection of chimeric plasmid containing colonies? 1) They're all equally good. 2) Hindll 3) EcoRI 4) BamHI

11. please fill in the blanks

Write TRUE or FALSE. If false, write the word/s that make(s) the statement incorrect. 1.The primosome complex includes primase, helicase and gyrase. 2.In plasmid DNA biotechnology, plasmid inserted with a “correct" DNA sequence is transferred to bacteria for rapid.

2) If the "isolated gene" from the above image happens to be the gene that codes for INSULIN, what will the bacteria be able to do when the gene is incorporated into its DNA?

VII. Match the technique described to the application given. DNA amplification b. РCR c. gel electrophoresis d. STR profiling dideoxy sequencing f. automated DNA sequencing g. site directed mutagenesis gene therapy h. 97. Nucleotides missing 3'-OH group makes small pieces that can be analyzed for identification. 98. DNA amplification without bacteria that includes cycling temperature changes to produce multiple copies of small pieces. 99. Identification of DNA by a series of peaks made from fluorescing based on fragment length. VIII. Match each item to the correct statement below. f. negative feedback system g. excretory system h. autonomic system i. positive feedback system j. parasympathetic nervous system homeostasis b. somatic system a. C. nervous system d. sympathetic nervous system endocrine system е. 100. way in which the body works to reverse a change and bring about normalcy 101. the tendency of the body to maintain a constant internal environment IX. Match each item to the…

4 I.I 5 | I6 I Use the following information to answer questions 14-15. A student has forgotten which antibiotic plasmid she used in her E. coli transformation. It could have been kanamycin, ampicillin, or tetracycline. She decides to make up a special set of plates to determine the type of antibiotic used. The plates below show the results of the test. Plate I. LB agar with kanamycin Plate II. LB agar with ampicillin Plate III. LB agar with tetracycline Plate IV. LB agar 14) Which antibiotic plasmid has been used? (kanamycin, ampicillin or tetracycline?) (a, b, c or d?) 15) Which of the following choices is best explanation for these results?

5. fill in the blanks please . You have identified 3 different alleles in human populations for part of the β-interferon gene that can easily be scored using PCR amplification followed by a restriction digest with XhoI:· Allele a1 has a 700-bp PCR product with no XhoI restriction sites.· Allele a2 also has a 700-bp PCR product, but due to a single-nucleotide polymorphism it has an XhoI restriction site 300 bp from one of the priming sites.· Allele a3 has a shorter 600-bp PCR product due to a deletion, and lacks an XhoI restriction site.You load the digested PCR products for three children (1, 2, and 3) along with their parents (M and F), in wells of an agarose gel (rectangles at the top of the diagram below), run the gel, and obtain the following results. The first lane contains a size marker with 100 bp increments; the bottom band is 100 bp long. For each individual, type in their diploid genotype, showing their two alleles using some combination of a1, a2, and/or a3, separating the…

The properties that all plasmids share are that they1. all carry genes for antimicrobial resistance.2. are self-transmissible to other bacteria.3. always occur in multiple copies in the cells.4. code for non-essential functions.5. replicate in the cells in which they are found.a) 1, 2    b) 2, 3    c) 3, 4    d) 4, 5    e) 1, 5

Select all the statements that provide evidence that DNA is the genetic material. Check All That Apply 1. By weight, a eukaryotic chromosome contains more protein than DNA. 2. Treatment of purified DNA with a DNA-degrading enzyme destroyed the ability of S bacteria to transform R bacteria. 3. The R mutant form of S. pneumonia does not cause infection in mice. 4. Radioactively-labeled DNA remained in bacterial cells after phage T2 infection. 5. Proteins are more complex than DNA because they are composed of 20 different amino acids. Pls help asap!! I am so confused