la: The 2x PCR Master mix has four main components. Two of these are PCR Buffer and MgC12. 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.
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- 3 Step3: After you have prepared the 3 diluted plasmid samples and the master mix, you can now use them to prepare the individual PCR reactions. For example, to prepare 25 μl PCR reactions for sample A, you need to combine, J ul of "diluted plasmid sample A", and M ul of "master mix". Repeat this step for all 3 reactions, then you can start the PCR reaction in the thermocycler. Solve for J and M.PCR is quick, efficient and easy to perform. However, there are some situations when cell-based cloning is preferred over PCR to amplify a DNA sequence. Mention two of them.Describe and contrast the common steps of DNA replication in vivo and the PCR reaction in vitro? In simple terms so, that I can understand. Thank you
- Are you a hidden heterozygote? A PCR analysis (part2) Agarose gel electrophoresis and interpretation la: Several factors (including agarose gel concentration, time and current) affect migration of DNA fragments through the agarose gel. Briefly explain how each of these factors affects DNA migration. Agarose gel concentration: Time: Voltage: 1b: Do DNA fragments move towards the positive or negative end of the gel box? Explain your answer. 1c: What is the purpose of the Tris-Acetate-EDTA (TAE) buffer that the agarose gel is prepared with and submerged in for running? What would happen if you used water to prepare and run the gel instead of TAE buffer? 1d: If the student is homozygous for the brown allele, how many bands will they see in the lanes for the blue and brown allele samples? (circle one) Brown sample: 0 Blue sample: 1 2 more than two. 1 2 more than two. le: If the student is homozygous for the blue allele, how many bands will they see in the lanes for the blue and brown allele…The process of DNA Replication depicted in the Polymerase Chain Reaction technology. Below are steps/components in DNA replication in cells. Briefly indicate the purpose/function of the components in DNA replication at the second column. For the third column, watch a PCR video in youtube and fill the column under PCR as to what step/components in the PCR technology corresponds to the second column. Question: In Reverse Transcription PCR, what step is added the ordinary PCR? Provide short explanation in the last column.PCR is a very useful method in biotechnology because it does not require the 6 or more different key proteins/enzymes required for DNA replication in living cells. Explain how and why this technique only requires 1 enzyme to make lots of DNA.
- Answer the following questions related to PCR Bioinformatics for DNA Extraction using Why is it necessary to chelate the metal ions from the solution during the boiling/lysis step at 100 degrees celsius? What would happen if you did not use a chelating agent such as the InstaGene matrix? What is needed from the cells for PCR? What structures must be broken to release the DNA from a cell?1. Explain what primers are and what purpose they serve in a PCR reaction. Explain the main steps and temperatures of one PCR cycle. 2. Explain the purposes of the following components of gel electrophoresis: agarose, loading buffer, DNA ladder/marker, electrode buffer, electrodes. 3. Explain the expected results of the Alu sequence at PV92 that we tested, in terms of numbers of bands and their sizes. A labeled drawing would suffice.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:
- show your rationale How many RT-PCR products generated from 13 copies of mRNA (= cDNA) from 33 PCR amplification cycles? What was the original starting number of mRNA molecules if after 36 PCR cycles, the reaction had generated 996,432,412,672 copies?. In the PCR process, if we assume that each cycle takes5 minutes, how manyfold amplification would be accomplished in 1 hour?Make the PCR Cocktail I field out my work.. but I don't know which one is incoeert.. This table lists the ingredients, stock reagent concentrations, and concentrations in the PCR reaction. Prepare a "PCR cocktail" to be added to your samples to achieve these concentrations. Make enough cocktail to run nine samples. [Four student samples + three positive controls + one negative control + one extra.] {Hint: Remember that the concentration in the reaction is not the same as the concentration in the cocktail!] Component Stock Concentration Concentration in the PCR reaction Volume per reaction Volume to make cocktail Sterile water - - 0µl 0µl PCR buffer w/ MgCl2 10x 1x 4µl 36µl Nucleotide mix 10 mM 0.2 mM 0.8µl 7.2µl Primer 1 (Forward) 10 µM 1.0 µM 4µl 36µl Primer 2 (Reverse) 10 µM 1.0 µM 4µl 36µl Taq DNA polymerase 5 U/µl 1.0 U 8µl 72µl DNA template (sample) - ~1 ng 20 µl 180µl Total - - 40 µl 360 µl