Q: COMPONENT 1 REACTION N+1 REACTIONS 15.80 µL 2.50 µL 1.50 µL 2.00 µL 1.00 µL 1.00 µL 0.15 µL…
A: Polymerase chain reaction is a procedure that is used to amplify a particular segment of DNA. This…
Q: QUESTION 3: Why does a PCR reaction require a primer? composed of DNA or RNA? Would you expect this…
A: The polymerase chain reaction, or PCR, is a chemical reaction used by molecular biologists to…
Q: The laboratory has lost the protocol to follow during the PCR reaction and only give you this.…
A: PCR or polymerase chain reaction is a method used for amplifying a small DNA sample. From a small…
Q: What is real-time PCR?
A: Besides tools, there a several techniques which are used in recombinant DNA technology. These…
Q: Recombinant pharmaceuticals (for the production of insulin, human growth hormone or blood clotting…
A: The human genome is composed of sequences of nucleic acids, which are encoded as DNA(Deoxyribo…
Q: Describe and contrast the common steps of DNA replication in vivo and the PCR reaction in vitro? In…
A: DNA replication and Polymerase Chain Reaction are both processes that create copies of DNA. DNA…
Q: What is experiment of Thermodynamics of Small Oligomeric Duplex DNA Denaturation.
A: Double-stranded DNA is the most common kind of DNA molecule found in nature. The strands can be…
Q: Why is it necessary to chelate the metal ions from the solution during the boiling/lysis step at 100…
A: The polymerase chain reaction (PCR) is a fundamental biochemical method for exponentially…
Q: ... and ..... are in product of a pcr reaction: primers nucleotides both none
A: Introduction :- PCR is described as the polymerase chain reaction . It is a bio-physical technique…
Q: You receive a tube containing 18.8 nmol of lyophilized (freeze dried) primers to be used in PCR. How…
A: Primers are a short nucleic acid sequence that provides a starting point for DNA synthesis. I…
Q: How do you know if it works? If you have more than one band in a lane, why what would be the reason
A: PCR is a polymerase chain reaction and it is used to amplify the segment of DNA/Nucleic acid.
Q: What are the importance of accurate wavelength measurement in purified DNA sample?
A: DNA is a double helical hereditary molecule. The nucleosome is a monomer of the DNA. In DNA, four…
Q: What are the roles of the following reagents in DNA extraction? a. Ethanol b. NaCl c. SDS d. TE…
A: A) The initial role of the ethanol and monovalent cations is to remove the solvation shell…
Q: Question 1. Restriction endonucleases can be isolated from a number of bacteria. In bacteria…
A: Restriction endonucleases are DNA cutting enzymes. They cleave the DNA at or near specific sequence…
Q: Question 1 During nucleic acid hybridization, the probe is labelled for DNA stability to increase…
A: Hybridization permits the distinguishing and cloning of explicit qualities, investigation of levels…
Q: Tell me how did you prepared the gel if the tray is 150 ml? how many milliliters of the TBE and how…
A: The answer to the first question is, 3 grams of agarose in 150 ml of TBE. *to get the second…
Q: Why different temperatures are used in a PCR reaction? Please answer at your own words.
A: The full form of PCR is polymerase chain reaction. Polymerase chain reaction is a process by which…
Q: Assume the samples in this diagram of gel electrophoresis are DNA molecules. Why do the samples…
A: Gel electrophoresis is used to separate molecules of different sizes. It uses charged molecules that…
Q: Recombinant pharmaceuticals (for the production of insulin, human growth hormone or blood clotting…
A: The recombinant DNA technology (cloning) is used for the production of insulin, growth hormone (GH),…
Q: Sequencing reactions are done in separate tubes for each ddNTP with a radioactive primer. Which…
A: In our Desire primer it is 15 nucleotide long however on gels there are only 10 bands are present…
Q: What is the first step that should be done in PCR testing?
A: PCR is the polymerase chain reaction. PCR test or polymerase chain reaction test is good method to…
Q: Our PCR samples already contain loading dye, but sometimes this isn’t the case. If your samples…
A: Gel electrophoresis is a process that helps in the separation of DNA fragments based on sizes with…
Q: Question 2. It is your first week working in the lab but unfortunately you program the PCR machine…
A: PCR is also called as polymerase chain reaction which has the ability to make millions to billions…
Q: What are the DNA extraction methods? Explain each one.
A: Ans: The Deoxyribonucleic acid (DNA) needs to be extracted out of cell for further studies on which…
Q: Please answer these two questions regarding PCR: a) Why do you need to perform PCR on DNA obtained…
A: The polymerase chain reaction (PCR) is a technique used to make many copies of a specific segment of…
Q: Why is it important to wear gloves when setting up the PCR tubes?
A: PCR represents polymerase chain reaction which is an in vitro laboratory technique used for the…
Q: QUESTION 5 Imagine that you have cloned the gene encoding the SARS-CoV-2 spike protein to a plasmid…
A: DNA cloning is the process of making multiple, identical copies of a particular piece of DNA. In DNA…
Q: When using a micro centrifuge, what should you check before starting the machine? A. That the…
A: Microcentrifuge is a type of laboratory centrifuge, which is used to separate particles in…
Q: QUESTION 9 Imagine that a researcher has a cell from a genetically engineered organism and a clone…
A: PCR or polymerase chain reaction is a widely used method for amplifying a particular DNA…
Q: During agarose gel electrophoresis, why does DNA move through the gel when electric current is…
A: Gel electrophoresis is basically a method that aid in separating DNA fragments and other molecules…
Q: The concentration of your RNA solution is 1500 ng/μl. How much RNA solution do you need to use when…
A: The concentration of RNA in solution can be determined by measuring absorbance at 360 nm.
Q: Question 1 A T What is the sequence of the sample DNA submitted for sequencing given the gel…
A: The unknown DNA samples can be sequenced by the Sanger sequencing method. This method is based on…
Q: The following image is of an agarose gel. If DNA samples were loaded to this gel and the…
A: Electrophoresis is a method that uses an electric field to separate macromolecules in a fluid or gel…
Q: Why does the temperature constantly fluctuate inside the PCR machine? What is the purpose of the…
A: PCR, or Polymerase Chain Reaction, is a biological technique invented by Kary Mullis.
Q: Question 5 Match the following components with the biotechnology technique they play a role in.…
A: Recombinant DNA technology is a technique by which foreign DNA is introduced into the host. DNA…
Q: how do you prepare 200 ml of 1.5 agarose gel in PCR?
A: Agarose is an uncharged, inert substance that does not bind to or react with molecules. Agarose is…
Q: What factors could explain a transformation efficiency?
A: Transformation efficiency is the efficiency with which a bacterial cell can take up the…
Q: What types of DNA can be extracted? Explain your answer.
A: Some of the most common DNA extraction methods include organic extraction, Chelelx Extraction and…
Q: QUESTION 4: Should PCR primers be complementary to each other?
A: Answer 4-:PCR primers should be complementary to the strands of DNA rather than complementary to…
Q: . Draw the melting curve for primer annealing, labeling the axes. Label the position of the…
A:
Q: What is the significance of using thermostable DNA polymerase for Polymerase Chain Reaction?
A: Polymerase chain reaction (PCR) is a technique for producing multiple copies of DNA in in-vitro…
Q: You set aside some of your purified PCR product to run on a gel. Name two things we learn by running…
A: Gel electrophoresis is a process that helps in the separation of DNA fragments with the help of an…
Q: what are the reagents and other equipment needed in polymerase chain reaction (or PCR)
A: PCR is a polymerase chain reaction which is used to amplify a given segment of DNA. PCR is a widely…
Q: Why are DNA samples that are to be separated by gel electrophoresis always loaded at the cathode end…
A: The method of electrophoresis is used to separate biomolecules based on particle charge, particle…
Q: What advantages does fluorescent labeling offer over radioactive methods of labeling DNA?
A: Fluorescent labeling and radioactive labeling have found wide applications in DNA sequence analysis…
Q: What is PCR? Why does Taq polymerase work better than a typical DNA Polymerase isolated from E. coli…
A: A groundbreaking move for molecular biology has been demonstrated by the discovery of polymerase…
Step by step
Solved in 2 steps
- Considering DNA sequencing by the Sanger method. It is correct to say that: * A)In the traditional method, radioactively “labeled” primers are used, allowing their visualization in autoradiography. B)In the automated method, a single reaction is performed containing the four “labeled” dideoxynucleotides, each with a different fluorophore. C)In both traditional and automated methods, the fragments are resolved and interpreted according to their ionization state. D)In the automated method, di-deoxynucleotides “labeled” with the same fluorophore are used, thus allowing their interpretation based on graphs of fluorescence emission. E)Sequencing reactions can use mRNA molecules, as long as they have a polyA tail.The optimal design of primers is critical to the effective amplification of DNA sequences. i) Outline the criteria for optimal primer design. i) Illustrate forward and reverse primer sequences using an example of a hypothetical DNA fragment. Give a brief overview of an online primer design software program.Draw a gel to represent the band shift assay result for testing the following mixtures: (1) Eukaryotic DNA along(2) Eukaryotic DNA + TFs(3) Eukaryotic DNA + RNA polymerase II(4) Eukaryotic DNA + TFs + RNA polymerase II(5) Eukaryotic DNA + TFs + RNA polymerase II + nucleotidesBands for smaller molecules can be ignored
- 2) The authors investigate if the switch from the polymerase to the exonuclease site involves releasing and re-binding of the DNA or if it utilizes an intramolecular rearrangement. To accomplish this, they use a primer-extension assay. How is a primer extension assay performed? In answering this question, be sure to mention a.) why heparin is used, b.) how they generate mismatched primers, and c.) how they visualize only the primer strand and not the template strand on their gel.What volume of 4X loading buffer must be added to 21 micro L of DNA in a technique for DNA sample preparation for agarose gel electrophoresis to generate a 1X buffer solution?1) What happened to the DNA at the different temperatures? How does Polymerase Chain Reaction exploit this property of DNA (Word count: 200) 2) Briefly outline the steps of DNA extraction as carried out in a lab for the purposes of sequencing, with reference to a published protocol from a company that supplies regents and kits for DNA extraction (e.g Qiagen, ThermoFischer, Invitrogen, or similar). Please Include the weblink for the published protocol you used as a reference
- 1) Prepare the following enzymatic reaction, present it in tabulated form. In a final volume of 30 ul, where buffer 4 (10 ml). How much volume of each reagent would be used and how much of water? Is there any problem? 2) The DNA pol 1 enzyme comes at a concentration of 50,000 U/ml. You have to prepare a 50 ug PCR reaction where you must use 0.05 U/ml reaction. You add 10 ul of PCR buffer, 2 ng of tempered DNA that is at a concentration of 0.5 ng/ul, primers (which are at 200 mM) so that each one remains at a concentration of 200 uM, Mg+2 that is 5 mM (10 X), enzyme and water. Present the table of all the reagents included in the reaction, the volumes of each one in ul. Present where the initial and final concentration of each reagent applies. Assume you have micropipettes for all values.ARE THEY TRUE OR FALSE? a) In a caesium chloride density gradient centrifugation method performed in the presence of ethidium bromide, supercoiled plasmid DNA binds more ethidium bromide than linear DNA. B)Tth DNA polymerase shows DNA-dependent DNA polymerase activity as well as RNA-dependent DNA polymerase activity. C)Magnesium ions stimulate polymerase activity. Therefore, it is better to use the highest concentration of magnesium in PCR amplification. D)When lambda bacteriophage infects E. coli cell lysis never occurs, and the infected bacterium can continue to grow and divide. E)The copy number refers to the number of molecules of an individual plasmid that are normally found in a single bacterial cell. F)RNase H selectively hydrolyzes phosphodiester bonds of RNA molecules in RNA:DNA duplexes.a)Dr. Thisisaneasyexam decides to amplify a gene from a plasmid using PCR. She starts out with 6.6 x 10-14g of a 10 kb template in a 100 µl reaction. Assuming that the molecular weight of the average base pair is 660 Daltons calculate the number of molecules of the template. b)Given that the concentration of each primer (20 base pairs each) is 0.1µM in this same reaction volume (100 µl) calculate the number of molecules of the primer present
- Below are 9 possible primer pairs. ● Determine which primer pair is the best choice by considering the following: 1. primers should be 18-24 bases in length; 2. base composition should be 45-55% (G+C); 3. primers should end (3') in a G or C, or CG or GC: this prevents "breathing" of ends and increases efficiency of priming; 4. Tms tween 55-70°℃ are preferred (Tas, annealing temperatures, are approximately 5°C lower than the Tm); 5. the Tm for your primer pair should be within 2 degrees of each other, though ideally the same; 6. runs of three or more Cs or Gs at the 3'-ends of primers may promote mispriming at G or C-rich sequences (because of stability of annealing), and should be avoided; 7. 3'-ends of primers should not be complementary (i.e. base pair), as otherwise the formation of primer dimers will result; 8. primer self-complementary (ability to form secondary structures such as hairpins) should be avoided. • Explain why the other primers are not good choices. ● Underline or…Answer the following parts: A. When performing classical Sanger or "dideoxy" sequencing, you set up 4 parallel reactions per template to be sequenced from a specific primer, with each of the four reactions containing a different dideoxynucleotide, and then the four reactions were run in a separate, adjacent lanes on a gel. Why couldn't you combine all 4 dideoxynucleotides with the primer and the template and do the whole reaction in one tube, and then run the set of fragments produced by the reaction mixture on a single lane in an acrylamide gel? B. When doing automated sequencing, on the other hand, all 4 dideoxynucleotides are added to the same sequencing reaction, and run together in a single capillary gel. What's the difference - why can an automated sequencing reaction be done with all 4 dideoxynucleotides mixed together and all the resulting fragments run together, but not a conventional sequencing reaction?It is desired to isolate genomic DNA from liquid culture of S. cerevisiae yeast. A commercial kit will be used to isolate genomic DNA from this liquid culture. Answer the following questions to understand the strategy used by commercial kits for genomic DNA isolation. a) List all the steps from cell pellet preparation to DNA elution. b) With which feature can the membrane in the column that comes with the commercial kit bind DNA? c) Which component in the kit would you use to recover the DNA from the membrane of the column to which the DNA was attached?