asap please. 3. A plasmid was cleaved with several restriction enzymes, individually and in combinations. The followingfragment sizes (base pairs) were determined by agarose gel electrophoresis.Eco RI4363Ava |2182Pvu ||4363Pst 14363Eco RI - Avo I2938Eco RIPst I3609Ava | - PvU II3722Avo1 - Pst |2182Pvu Il-Pst |2820Eco RIPvu II 2297142575464115432066Make a restriction map based on this data.Note: There may be some slight discrepancy in summing up the total base pairs. Indicate the distances be-tween sites. Why is only one band detected in the Ava 1 - Pst I co-digest?

Plasmids include genetic resources, and each plasmid has an attachment site, an identification site,…

A plasmid was cleaved with several restriction enzymes, individually and in combinations. The following fragment sizes (base pairs) were determined by agarose gel electrophoresis. Eco RI Aval Pvu || Pst | 4363 2182 4363 4363 2938 3609 3722 2182 Eco RI - Avo I Eco RI - Pst I Ava | - Pvu II Aval - Pst | Pvu II - Pst I 2820 Eco RI - Pvu II 2297 Make a restriction map based on this data. Note: There may be some slight discrepancy in summing up the total base pairs. Indicate the distances be- tween sites. Why is only one band detected in the Avo I - Pst I co-digest? 1425 754 641 1543 2066

A plasmid was cleaved with several restriction enzymes, individually and in combinations. The following fragment sizes (base pairs) were determined by agarose gel electrophoresis. Eco RI Aval Pvu || Pst | 4363 2182 4363 4363 2938 3609 3722 2182 Eco RI - Avo I Eco RI - Pst I Ava | - Pvu II Aval - Pst | Pvu II - Pst I 2820 Eco RI - Pvu II 2297 Make a restriction map based on this data. Note: There may be some slight discrepancy in summing up the total base pairs. Indicate the distances be- tween sites. Why is only one band detected in the Avo I - Pst I co-digest? 1425 754 641 1543 2066

Biology: The Unity and Diversity of Life (MindTap Course List)

15th Edition

ISBN:9781337408332

Author:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr

Publisher:Cecie Starr, Ralph Taggart, Christine Evers, Lisa Starr

Chapter15: Studying And Manipulating Genomes

Section: Chapter Questions

Problem 16SQ: Match the method with the appropriate enzyme. _____ PCR a. Taq polymerase _____ cutting DNA b. DNA...

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Match the method with the appropriate enzyme. _____ PCR a. Taq polymerase _____ cutting DNA b. DNA ligase _____ cDNA synthesis c. reverse transcriptase _____ DNA sequencing d. restriction enzyme _____ pasting DNA e. DNA polymerase (not Taq)
1. Briefly explain why the total size of the pMBBS plasmid in the Restriction Enzymes practical is 3000bp (base pairs) 2. Briefly explain why the cut sites on the pMBBS plasmid in each Restriction Enzymes (EcoRI, BamHI, and XhoI) just 1 each 3. Briefly explain what led to the 5 fragments formed which linked to the 500bp, 1000bp, 1500bp, 2000bp, 2500bp sizes
1. Given the following restriction endonucleases and the sequences of their corresponding restriction sites, which would be LEAST useful for cutting the plasmid and the foreign DNA to be inserted? [The arrows indicate where cuts are made.] (see img) a. EcoRI b. HindIII c. HaeIII d. BamHI 2. EcoRI and HindIII are two different restriction enzymes. If the DNA from two different sources were cut with either EcoRI or HindIII, which of the cut DNA fragments would NOT join together easily so that they could be sealed with ligase? a. E. coli DNA cut with EcoRI and human DNA cut with HindIII b. E. coli DNA cut with HindIII and mouse DNA cut with HindIII c. human DNA cut with EcoRI and chimpanzee DNA cut with EcoRI d. mouse liver DNA cut with HindIII and mouse kidney DNA cut with HindIII
The following data is from the analysis of a circular piece of DNA (plasmid). A. Use this data to generate a circular map of the plasmid. Pst I: 10 kb Bam HI: 10 kb Sal I: 4.5, 3, 2.5 Sal I + Pst I: 4.5, 3, 2, 0.5 Pst I + Bam HI: 7, 3 Sal I + Bam HI: 4.5, 2.5, 2, 1 B. You have purified the 3 kb Pst I to Bam HI fragment of DNA and have labelled the DNA for use in a Southern blot. If the double cuts are shown were run on a gel, transferred to nitrocellulose, and then analyzed by probe hybridization; which band or bands from each double-cut would be identified?
1. Scientists were able to develop transgenic bacteria that can be used for mercury remediation. This process involved combining genes for mercury resistance (so that the bacteria itself has protection) and mercury accumulation (thereby allowing the bacteria to sequester Hg for retrieval later on). They probably used the following techniques in creating the transgenic bacteria EXCEPT a. plasmids b. DNA ligase c. DNA polymerase d. restriction enzymes 2. Consider these two parents (Mom and Dad) and their son, Calvin. (see pic attached). Which two members of this family would show the greatest difference in their DNA profile?
A student wanted to load .75 ug dna on agarose gel and has 4x loading buffer for sample preparation. The student has 50ul purified plasmid, finding concentration of plasmid to be 250 ng/uL The student used 10uL plasmid DNA in 50uL reaction for the restriction digest Give the volumes of restriction digest and 4x loading buffer that student would mix together and load on agarose gel.
1. You have the plasmid pUC18/19, which is a circular plasmid that consists of 2686 bp. What would the number of and length of the fragments be if you cut the plasmid with the following restriction enzymes or combination of enzymes? Give a schematic representation of the digestions.a. PscI & GsuIb. ScaI, PdmI & BsaXI c.ScaI, SspI & EheI
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.
1. a) A technologist has a 20µl sample of DNA and adds 5µl of loading dye before adding the total volume into an agarose gel. What was the original concentration of the loading dye? a. 4x b. 5x c. 6x d. 7x e. None of the above 1. b)A technologist adds 6uL of 5x loading dye to a DNA sample before adding the total volume into the agarose gel. How much DNA sample was combined with the loading dye? 12uL 16uL 20uL 24uL 28uL
Margaret has been given a plasmid containing her favorite gene, afg (Margaret’s favorite gene). The only thing sheknows is that afg was cloned into the vector using a single restriction enzyme, BstBI. Margaret orders theBstBI enzyme from NEB, so she can cut the plasmid and confirm the presence of the insert. On theNanodrop, she measures the concentration of her plasmid to be 401 ng/μL. Describe how she would setup and perform this reaction if she wanted to digest 2.0 μg of the plasmid.
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+ HindlII+ Pstl. label on the plasmid. i) The location of the Pstl site ii) The sizes of the new fragments made by this Pstl cut
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)