For questions 16-19, please use the word bank in the box below to fill in the sp
ID: 149746 • Letter: F
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For questions 16-19, please use the word bank in the box below to fill in the spaces. mRNA, RISC, cDNA, gDNA, PCR. RNAase-H, drosha, viral, non-viral, hydrogen, 55oC, 72oC, 95oC, two- fold, PCR, reverse transcription, primer, reverse transcriptase, Poly-A tail, uciferase, dNTP, positive negatively, RNA Polymerase, ATP, dicer, guide RNA, endocytosis, ATP sulfurylase, apyrase, pyrophosphate, luciferase APS, luciferin, 16. Below is the description of method used for gene expression analysis using RT-PCR. Filli the blank. (15pt) "The ability to synthesize DNA from an RNA template, via enables researchers to study is generated by the which has the ability to use the information in an RNA to generate a is a RNA-dependent DNA polymerase. Like all DNA RNA with the same molecular approaches used for DNA investigations. enzyme complementary DNA. Thus, polymerases it cannot initiate synthesis de novo but depends on the presence of aSince many mRNAs have a initial cDNA has been generated it is generally necessary to produce a second strand of DNA. at the 3' end, oligo-dT is frequently used to prime DNA synthesis. Once the has the ability to cause single-stranded nicks in the RNA, and DNA polymerase can then use generated by reverse , which allows the detection of low abundance RNAs in is denatured by heating to which bonds between complementary strands, yielding single-stranded molecules. The in order to allow primers complementary to the sequence(s) of these single-stranded nicks to initiate " second strand" DNA synthesis. transcription can be amplified using a sample. In the first step of the PCR process, the disrupts the temperature is then lowered to interest to anneal. The DNA polymerase included in the reaction will then begin DNA synthesis. At this point, the temperature is raised to the optimal activity temperature of the DNA polymerase at synthesize a new strand complementary to the template. The process of denaturing, annealing, and extension can be repeated multiple times, witha Because PCR can selectively amplify a template, it is an important method for detecting specific nucleic acid molecules in a particular cell or small populations of cells." to increase in the amount of DNA molecules with each cycle 17. Below is the description on the mechanism of RNA interference. Fill in the blank. (7pt) transcribes the pri-miRNA in the nucleus that forms a stem-loop. Then, cleaves the pri-miRNA at two sites to create a 65-70-nucleotide-long pre-miRNA. When it is transported to the cytoplasm, The double-stranded RNA is denatured to become the single stranded into the Then, the through slicer by clearing the target mRNA." cleaves the loop from the pre-miRNA to a 22-nucleotide-long double-stranded RNA that can be incorporated brings the target mRNA, which can repress gene expressionExplanation / Answer
Q.16
1. Reverse transcription [Explanation: Synthesis of cDNA from mRNA is known as reverse transcription]
2. cDNA [Explanation: It is known as complementary DNA which is synthesized from mRNA]
3. Reverse transcriptase [Explanation: It is an enzyme which catalyse the reverse transcription]
4. Reverse transcriptase [Explanation: It is an enzyme which catalyse the reverse transcription]
5. Primer [Explanation: It is a short stretch of sequence which is used during the initiation of DNA synthesis]
6. Poly-A tail [Explanation: Found at 3’ end of mRNA sequence]
7. RNAase-H [Explanation: Degrades the RNA molecule after cDNA synthesis]
8. gDNA [Explanation: It’s a double stranded DNA molecule ]
9. PCR [Explanation: Used to amplify the gene of interest]
10. gDNA [Explanation: It’s a double stranded DNA molecule]
11. 950C [Explanation: This temperature will denature the gDNA]
12. Hydrogen [Explanation: DNA strands are held together by hydrogen bonds]
13. 550C [Explanation: Primer gets annealed to the DNA strand at this temperature and it should be lower than extension temperature]
14. 720C [Explanation: Extension of DNA happens at this temperature and it should be higher than annealing temperature]
15. Two-fold [Explanation: After every cycle of PCR amplification, we can observe two-fold increase in the template concentration ]
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