Compare Illumina/Pacbio/Nanopore sequencing methods. 2. How do you construct the profile matrix? Answer all the part in details and with proper examples.
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1. Compare Illumina/Pacbio/Nanopore sequencing methods.
2. How do you construct the profile matrix?
Answer all the part in details and with proper examples.
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- explain how the following genotypic andphenotypic methods of microbe identification is done. Use diagrams or schematic representations to explain yourwork. 1. Pulsed-Field Gel Electrophoresis (PFGE)2. Whole Genome Sequencing (WGS)1. How does PFGE separate larger fragments more efficiently than standard electrophoresis? 2. Why is SYBR green less toxic than EtBr? 3. What are the similarities and differences between Manual and Automated Sanger Sequencing? 4. What is the relationship between DNA fragment length and the distance it will run in a gel? (Restriction Enzyme Digestion)1. What are the things to consider in making a good primer? 2. Does temperature affects the primer design? Discuss briefly.
- 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?What do you think are the crucial steps in Polymerase Chain Reaction technique? Enumerate the advantage and disadvantage of Polymerase Chain Reaction technique? What are the things to consider in making a good primer? Does temperature affects the primer design? Discuss briefly.Discuss the principles , uses, advantages and disadvantages of illumina sequencing method
- what is the |usage of DNA separation in gel electrophoresis please write the resources under the answerDiscuss the use of gel electrophoresis for the separation of macromolecules (DNA, RNA and protein) of different sizes and topological forms. (Include in you answer the different recombinant DNA techniques that require gel electrophoresis)1. You decide to perform dideoxy sequencing on a PCR product. You add the appropriate 32P- labeled primer, DNA polymerase, DNA template (the PCR product), buffer, DNTP mix, and a small amount of one of the four ddNTPs to four reaction tubes. You run the reactions in the thermal cycler, load each reaction into a separate lane of a polyacrylamide gel, and separate the products by gel electrophoresis. In the figure below, the lanes are labeled according to the ddNTP added. ddATP ddTTP ddCTP ddGTP - Lane 1 2 3 4 a) In lane 5, draw what you would expect to see if you prepared a reaction using a nucleotide mix containing only DATP, DTTP, dCIP, and dGTP. b) In lane 6, draw what you would expect to see if you prepared a reaction using a nucleotide mix containing only ddATP, DTTP, dCIP, dGTP. ||
- 1. Definitions and brief statements 1. DNA Data Bank of Japan 2. MeSH Database 3. Please describe the 3 steps and temperature that are cycled during a PCR reaction. 4. What are global alignment and local alignment and their algorithms? ( 5. ProtScaleSequencing reactions are done in separate tubes for each ddNTP with a radioactive primer. Which picture shows the correct 4 reactions after separation of the sequencing reaction by gel electrophoresis? Template: Primer: 5'-ATCGCTTACCATTAG-3' 5'-CTAAT-3' ddA ddC ddG ddT ddA ddC ddG ddT = D ddA ddC ddG ddT A ddA ddC ddG ddT — B C4. Create a PCR master mix for the following: 48 samples, 25 uL total volume per reaction, 2 uL template DNA volume, 2.5mM MgCl, 0.5 µM each primer. Your stock concentrations are: reagent mix (2X, does not contain MgCl,), primers (F/R; 10 uM), MgCl, (50 mM). The master mix calculation must include a 10% overage, and all answers are to be given to one decimal place.