The Centers for Disease Control and Prevention (CDC) programPulseNet usesgenomic fragments that have been generated by restrictionendonucleasesO pathogen specific primersO PCR combined with restriction fragment length polymorphism (RFLP)analysisO oligonucleotide probes in microarray technology

Pulsenet uses highly multiplexed amplicon sequencing (HMAs) panels for surveillance subtyping. It…

Answered: The Centers for Disease Control and…

BIOLOGY:CONCEPTS+APPL.(LOOSELEAF)

10th Edition

ISBN:9781305967359

Author:STARR

Publisher:STARR

Chapter15: Biotechnology

Section: Chapter Questions

Problem 8SA: PCR can be used ______. a. as a cloning vector b. in DNA profiling c. to modify a human genome

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What type of probe is used for real-time PCR? Explain howthe level of fluorescence correlates with the amount of PCRproduct.
All of the following are performed during restriction fragment length polymorphism analysis. 1. splitting of double-stranded into single-stranded DNA 2. gel electrophoresis 3. autoradiography 4. immersion in radioactive probes 5. digestion of DNA with restriction endonucleases 6. use of a positive charge to transfer single-stranded DNA from a gel to a membrane. The correct sequence of these operations is what
Besides real time PCR, elaborate on three (3) other PCR techniques available through modification of the early PCR system.
You wish to insert a gene in the plasmid below. (Note the neon green lines represent restriction sites.) amp gene BamHI Sacl Saçl EcoRI BamHI 33 BamHI Gene of interest Chromosomal DNA from human cells a) Which restriction enzyme should you use to cleave the plasmid? Explain your answer. b) How would you create a cDNA library for the chromosomal DNA containing the gene of interest? c) The green area on the plasmid is the ampicillin resistance gene. Explain how you would use it to screen for a recombinant plasmid. 1
Which of the following scenarios would ONLY occur if your skipped the digest purification step? UV/VIS spectrophotometric quantification of DNA may be skewed by uncut plasmid. Some plasmid molecules may be cut once, by one enzyme, and re-ligate to themselves. The fragments cleaved by the restriction enzymes on the plasmid and insert can re-ligate to their sites, causing reduced ligation efficiency. Plasmid molecules cut with EcoRI and Xbal can ligate to each other instead of the insert.
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In the following "gene library" cloning experiment Digested genomic DNA AmpR gene TCR gene TCR is tetracycline resistant marker, AmpR is ampicillin resistant marker and BamHI is the unique restriction enzyme on plasmid. A PhD student digests/cuts the plasmids with BamHI restriction enzyme and the genomic DNA with EcoRI restriction enzyme. After performing the cloning experiment and obtaining colonies on a selection plate, the obtained cells will be ..... (Hint: this question is even more challenging; the PhD student was later demoted to an MSc student). a) resistant to ampicillin and tetracycline b) sensitive to tetracycline and ampicillin c) resistant to tetracycline and sensitive to ampicillin d) resistant to ampicillin and sensitive to tetracycline e) sensitive to ampicillin and tetracycline BamHI
A result of agarose gel electrophoresis of plasmid DNA is shown below. The direction of the electrophoresis is indicated by the arrow head. Lane 1 shows the plasmid DNA with no restriction enzyme digestion. Lane 2 shows the plasmid DNA which has been digested by restriction enzyme ECORI. Given the fact that there are only relaxed and supercoiled forms existing in this plasmid, please annotate the bands in Lane 1 on this gel with their supercoiling status (supercoiled or relaxed) and explain why. Then based on the information from Lane 1 and Lane 2, analyze the number of EcoRI cleavage sites on this plasmid. Lane 1 2
Using a ThermoFisher GeneJet Miniprep plasmid isolation kit to isolate plasmids from bacteria: Use of RNAse increases purity of purified plasmids. This enzyme will be added when the cells are lysed, detroying all RNA in the sample. Why will this action increase the purity of an isolated plasmid sample? This will only be used for the plasmids isolated using the GeneJet purification columns, and not when performing alkaline lysis. How will this affect the predicted results for the alkaline lysis plasmid isolations?
As you learned in this chapter,restriction enzymes are sophisticated“scissors” that geneticistsand molecular biologists routinelyuse to cut DNA for recombinant DNAexperiments. A wide variety of onlinetools assist scientists working with restrictionenzymes and manipulating recombinantDNA for different applications. Herewe explore Webcutter and Primer3, twosites that make recombinant DNA experimentsmuch easier.Exercise I – Creating a RestrictionMap in WebcutterSuppose you had cloned and sequenceda gene and you wanted to design a probeapproximately 600 bp long that could beused to analyze expression of this gene indifferent human tissues by Northern blotanalysis. Internet sites such as Webcuttermake it relatively easy to design experimentsfor manipulating recombinant DNA.In this exercise, you will use Webcutter tocreate a restriction map of human DNAwith the enzymes EcoRI, BamHI, and PstI.1. Access Webcutter at http://www.firstmarket.com/cutter/cut2.html.Go to the Study Area for…
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