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- What is the final volume of the individual PCR reactions we are making?
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- B. Restriction Mapping. Single and double digestion of plasmid pMCS326 were performed using the restriction enzymes Alulll and EcoRV. DNA fragments are shown in an electrophoretogram below. Construct a restriction map of plasmid pMCS326 for enzymes Alulll and EcoRV. 20 kb 11 kb 8 kb 6 kb kb 3 Alulll + Alull EcoRV ECORV | || Restriction Map:. In the PCR process, if we assume that each cycle takes5 minutes, how manyfold amplification would be accomplished in 1 hour?After transduction we move to mutagenesis analysis. Using the transduction plate,PCR I found that lacZ,fliE,nagC,ompA all show successful transduction and successful PCR. But for DNA sequencing some clonies are not correct. Expected size of PCR are lacZ 304,fliE 255,rpoA 420,nagC 329,ompA 731,lexA 350,kan cassette 549,tet cassette 265. Please indicate which mutation were successful and success why some were not from the size about. indicate at what stage any of the mutagenesis failed.
- 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 + |OO HUAWEI Nova 2 Plus DUAL CAMERA estion To study the function of any gene of interest you would perform the loss and gain of function approaches by either deleting or re-expressing the gene of interest, which of the following can be used to determine and quantify the activity of the gene? red Oa. Microscopy d out of O b. Western blotting O C. PCR/OLA on O d. Gene knockdown O e. DNA hybridization stion Which one of the following is NOT correct regarding Bacterial Biosensors? O a. produced by all Pseudomonas species ed O b. Encoded by LacZ gene out of O c. Encoded by Lux genes O d. Chemical compounds used for the quantitative assessment of water pollution O e. In the presence of pollutants the bioluminescent decreases. stion CD4-Pseudomonas Exotoxin Fusion Protein is another biotechnology strategy for HIV therapeutic setting. This construct will target and kill: Select one: d O a. Any lymphocyte put of O b. HIV (virus only) O C. Non infected TH O d. Infected TH cells O e. Infected TH…PCR Analysis la: The 2x PCR Master mix has four main components. Two of these are PCR Buffer and MgCl2. Based on your knowledge of DNA replication and PCR, what are the other two components and what are their functions? 1b: Explain what would happen if the PCR reaction did not contain Taq polymerase? Assuming all other required PCR components are present, would any part of the PCR reaction occur? Explain your reasoning. 1c: Why does a PCR reaction require a primer? Would you expect this primer to be composed of DNA or RNA? Explain your reasoning. 1d: Should PCR primers be complementary to each other? Explain your reasoning le: As you know, experiments include controls. In this lab, a negative control was included along with your experimental samples. Which components do you think the negative control would contain? Hint: Think about which component would be left out of the negative control relative to the experimental samples. Explain why. 1f: What would the PCR product be for the…
- O e. Gene amplification CLEAR MY CHOICE To study the function of any gene of interest you would perform the loss and gain of function approaches by either deleting or re-expressing the gene of interest, which of the following can be used to determine and quantify the activity of the gene? Oa. Western blotting O b. Gene knockdown O. PCR/OLA O d. Microscopy O e. DNA hybridization Paternity testing can be detected most precisely by using techniqueCloned Libraries You are running a PCR to generate copies of a fragment of the cystic fibrosis (CF) gene. Beginning with two copies at the start, how much of an amplification of this fragment will be present after six cycles in the PCR machine?Please answer this asap. Thanks, You have discovered a new plasmid RK21 in a unique bacterial community. As a first step towardunderstanding this plasmid, you digest the plasmid with three restriction enzymes: SspI, XhoI andSmaI. You run the digested plasmid DNA on an agarose gel, along with an uncut sample of theRK21 plasmid DNA as a control.Unfortunately you forget to load a DNA ladder, and obtain the following results. Assumecomplete digestion of all samples or all the digests worked completely
- Describe PCR. Give a real world example of when PCR may be used in the lab to solve a problem. Do a little research if nothing comes into your mind. For the toolbar, press ALT+F10 (PC) or ALT+FN+F10 (Mac). B I US Paragraph Arial x² X₂ ¶ Ⓡ ||| ||| 冈 A v 880 V Ix % $1 田田田由用Part A. If student counts 63 colonies on their 10^-6 dilution LB plate. What was the original concentration of their cells if they plated 100ul? Part B.If we used pGFPuv as the template for PCR positive control. This is because: a. it contains the GFP gene so it should show a product. b. It contains DNA fragments that were added to the ligation reaction. c. It is the desired plasmid we wanted to make. d. So we have a band to compare our unknown plasid to allowing us to check if the unknown is the right size..How much PCR product (500 bp) should be added to a ligation in which 50 ng of Vector (6.6 kb) vector will be used if a 3:1 insert:vector molar ratio is desired? Vector concentration is 50 ng/uL and insert concentration is 35 ng/ul. Provide mass and volumes for the ligation reaction