notes 15 quiz 15

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

1.  Below is an illustration of two plasmids: the incomplete pKAN-R which contains the gene of interest (rfp), and the good plasmid vector pARA which contains the ampR gene for ampicillin resistance, the araC activator gene and the ori region. Based on this, how are you going assemble to create the desired recombinant plasmid containing all the necessary components? (Keywords: Identify correct restriction enzymes to use to create specific sticky end sequences that will only assemble with the final vector in one orientation)

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Q. What is the total size of your recombinant plasmid vector containing the insert?   give answer asap

Second part asks: Explain the roles two of those enzymes would play in obtaining the recombinant plasmids

Answer choices for A: deletion, transversion, insertion, transition Answer choices for B: nonsense, framshift, silent, in-frame, missense Answer choices for C: mismatch repair, homologous recombination repair, nucleotide excision repair, base excision repair, photo reactivation repair, translesion snythesis, nonhomologous end joining Answer choices for D: conditional, loss-of-function, neutral, gain-of-function, lethal Answer choices for E: bottom, top Answer choices for F: 5' UTR, exons, stop codon, 3' UTR, introns

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Which of the delivery techniques mentioned in Table 2 can be used to best transfer recombinant plasmids into E. coli cells

What are DNA probes  Describe three (3) methods that can be used to label DNA probes  Describe 2 methods for detection of the hybridized probe List the advantages of using non-radioactively labeled probes vs radioactively labeled probes.

Q. Based on your understanding of plasmids’ answer the following questions.1. Why is a daughter cell that fails to inherit a kind of plasmid dies? 2. How transfer of Col E1 plasmid to an F cell occurs? 3. What component you will must include in constructing a plasmid cloning vector that can replicate in both bacteria and yeast? 4. How copy number of a plasmid may affect its segregation? 5. How the yeast 2u circle maintains 80 copies in a cell?

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Primers designing for epitope tagging: Design forward and reverse primers to amplify the following gene with 6×HIS-tag on the N-terminus of the protein. To be cleaved and inserted into the plasmid, add restriction sites for EcoRI and HindIII at 5' and 3'. ATGCTCTCCGCCCTCGCCCGGCCTGTCAGCGCTGCTCTCCGCCGCAGCTTCAGCACCTCAGCC CAGAACAATGCTAAAGTAGCTGTGCTAGGGGCCTCTGGAGGCATCGGGCAGCCACTTTCAC TTCTCCTGAAGAACAGCCCCTTGGTGAGCCGCCTGACCCTCTATGATATCGCGCACACACCC GGAGTGGCCGCAGATCTGAGCCACATCGAGACCAAAGCCGCTGTGAAAGGCTACCTCGGAC CTGAACAGCTGCCTGACTGCCTGAAAGGTTGTGATGTGTAA

A.How could endonucleases interfere with the transformation procedure? B. Does supercoiled or nicked plasmid get transformed more efficiently? Why?

Clone number in this case is number 196 which is shown in the images.  State whether a BamHI site has been re-created at the forward- and the reverse-end junctions of the human DNA with the plasmid vector  band sizes are shown in one of the images. (0.5, 1, 2, 3, 4, 5, 6, 8, 10kb)

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...]

1. How large is your plasmid 2. Which colony(-ies) is(are) likely to have been white? 3. Which colony(-ies) is (are) likely to have the XRAY gene?    asap please

33) Judging by mosaic clone images above, what effect does loss of dmyc likely have on cell growth? Please briefly explain.

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KINDLY PROVIDE 2 SIGNIFICANT DIFFERENCES EACH TERMS

please explain topic is about how  recombinant DNA is  made

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…

Kha Vu Danels Include: 8DX : Safehy Jor bromie tnto lab repart! Name Section date sheet MAPPING PRACTICE #1 Below is a restriction map for the plasmid PGEN 101 (total length = 20 Kb). Using this map as a guide, give the number of restriction fraqments along with their associated lengths that would result from digesting PGEN 101 with the restriction enzymes EcoRI, BamHI and a combination of ECORI and BamHI. BamHI 3.2 Kb 1.7 Kb EcoRI BamHI PGEN 101 8.7 Kb 5.5 Kb .9 Kb EcoRI ECORI DIGESTION PERFORMED SIZES OF FRAGMENTS OBTAINED 10.4 kb , 0.9kb, 8.7 Kb EcoRI 3.2 Kb, 16. 8kb BamHI EcoRI + BamHI

Explain your experimental approach if you must correct the genetic defect phenylketonuria, which is caused by mutations in the phenylalanine hydroxylase gene using the CRISPR/Cas9 system and homology directed DNA repair.

Using the figure below briefly explain how recombinant lentiviral particle can be formed.

7. Consider the 3 mechanisms of HGT in prokaryotes and identify 1) which one leads to the presence of a spacer in the CRISPR locus of a bacterial chromosome and 2) describe how this HGT mechanism occurs. Hint: in 2) the question asks to describe the HGT mechanism, not how the spacer is incorporated into the bacterial chromosome by Cas proteins. The HGT mechanism that leads to a spacer in a bacterial chromosome is During this mechanism

PCHEM4321. An agarose gel electrophoresis pattern of the plasmid PSPM4321 digestion (restriction) is shown below. Draw a restriction map of a plasmid with the appropriate restriction sites based on the data given below. Hindlll Hindll BamHI +BamHI Figure 1: 1% agarose gel electrophoresis of pCHEM4321 40 24 16 12 12 8 4 4 + |

Explain:a. functionalization of a sensor array with DNA oligonucleotide capture probes for ribosomalRNA (rRNA) species-specific sequences.b. sandwich hybridization of target rRNA with the capture probe and a horseradish peroxidase-linked DNA oligonucleotide detector probe.

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