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- 4. Explain the mechanisms of SYBR green and TaqMan in real-time PCR. 5. Explain what Ct mean and what it tells us about our samples in real-time PCR. 6. Explain why a positive test for COVID19 would appear sooner than a negative result when using real- time PCR to test if someone is infected.3. Match the following biotech tools with their applications. Some of those tools may have more than one applications. • A. Gene cloning and Recombinant DNA • B. Restriction enzyme • C. Radioactive nucleic acid probe • D. Reverse transcriptase • E. Ti plasmid. • F. Gene therapy • G. PCR • H. Gel electrophoresis • I. single nucleotide polymorphism (SNP) and Restriction fragment length polymorphism (RFLP) • J. Genomic 1. Crime scenes and paternity 2. Making copies of DNA sequence 3. GMO 4. introduce new genes into plant cells 5. alteration of an individual's DNA to treat disease. 6. Cutting DNA at target sequence 7. Detect the presence of specific gene 8. produce a DNA strand from MRNA. 9. separate DNA molecules based on size 10. methods to get the DNA fingerprint of different individual1. What are the reaction components, and what equipment do you need for PCR?2. What can be possible source of DNA for PCR?3. What are some uses for PCR?
- 18. If you tried to run a PCR with only three of the four dNTPS (GATP, dCTP, dGTP, but no dTTP) The reaction will work fine The reaction will work, but will be slower The reaction will not work because it needs ATP, not dATP, to provide energy to drive the reaction The reaction will not work because all 4 dNTPs are required to synthesize DNA The reaction will work if there is no Cytosine in the template PCR product after the second thermo1. What does PCR stand for? I 2. Explain the main objective of the PCR technique.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.
- 4. state why simplicity and speed are top priorities in selecting DNA isolation method4. 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.2. What components do you need to perform PCR? 3. What is in the master mix and why do you need each component? 4. Why do you need to perform PCR on DNA evidence from a crime scene? 5. What steps make up a PCR cycle, and what happens at each step? 176 Introduction to PCR-Based Forensics
- 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. What was the purpose of the clean-up pcr? How to know if it Was the clean-up successful?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.