DNA extraction, PCR, gel electrophoresis, and DNA sequencing of PCR products. Give a description of these methods and the materials required to perform each of these molecular techniques
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DNA extraction, PCR, gel electrophoresis, and DNA sequencing of PCR products. Give a description of these methods and the materials required to perform each of these molecular techniques
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- DNA Profiles as Tools for Identification A PCR-based paternity test is conducted using STRs that consistently produce a unique DNA fragment pattern from a single chromosome. Examining the results of the following Southern blot, which male(s) can be excluded as the father of the child? Which male(s) could be the father of the child?Briefly describe what happens during each of the phases of PCR (denaturation, annealing, and extension), including when you need each of the major components of the reaction (template, primers, nucleotides, DNA polymerase).Microarray hybridization is used mostly in transcript profiling or assaying DNA variation. Although the technology for establishing DNA microarrays was developed only recently, numerous applications have already been developed and their impact on future biomedical research and diagnostic approaches is expected to be profound. Give some examples of the practical use of this technique.
- 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 supplying reagents and kits, noting the refinements compared to the protocol used in the online practical. Include the weblink you used.The current state-of-the-art in forensic DNA profiling involves the PCR-amplification and analysis ofshort tandem repeats, STRs, in the human genome. This approach has many distinct advantages.Please list and explain three of those advantages.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 supplying reagents and kits (eg. ThermoFisher, Qiagen, Invitrogen or similar), noting the refinements compared to the protocol used in the online practical. Include the weblink you used.
- using the process flow of PCR describe the principle associated with the PCR technique.Discuss the principles , uses, advantages and disadvantages of illumina sequencing methodAnswer 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?
- PCR is a powerful technique to screen and amplify segments of DNA for use in recombinant protein technologies. Describe in detail, the components of a PCR reaction and why they are requiredBriefly 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). You should note the refinements compared to the protocol used in your practical session.Transcriptome analysis involves two separate methodologies: gene expression and RNA seq analyses. The 10 items below are a scrambled listing of the steps used in the two procedures. Identify the steps involved in RNA seq from the list below. Use the numbers in the list to refer to each step. Once the steps for RNA seq have been identified, write the steps in the order in which they are performed during the experiment. (1) DNA sequencing (2) Allow for hybridization and wash excess cRNA. (3) Mix labeled cRNA with array chip. (4) PCR amplification (5) Measure fluorescence intensity to determine abundance of transcripts. (6) Add labeled cRNA at each microarray location. (7) Map cDNA sequences to the genome of the organism to determine identity and abundance of transcripts. (8) mRNA isolation from cells (9) Prepare fluorescently labeled cRNA probes (10) cDNA synthesis