Tee - Lab_Report_DNA_RNA_Central_Dogma

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DNA, RNA, and the Central Dogma Tee Nguyen 4/25/23
Instructions: 1. Please read all of the introduction and background information within the investigative manual. a. Once you have done so answer the prelab question BEFORE completing any of the lab’s activities. 2. Once you have completed the prelab questions proceed to the activities of the lab within the investigative manual. a. As you read through the instructions for completing each activity make sure you also: i. Complete any instructions (append photos, etc)/ and answer any questions found in the post lab questions for each activity. ii. Take the photos of your experiments in each activity as directed below. IMPORTANT: Don’t clean-up your lab until you know what portion of the experiment you need to take a picture of. 3. Here is a video that will introduce you to the lab and its main concepts. The student is encouraged to watch it. a. DNA, RNA, and the Central Dogma Prelab Questions 1. In the investigative manual the processes of DNA replication, transcription and translation were described. While the descriptions are accurate and helpful videos of these processes are also very helpful. Please watch the following videos and then answer the prelab questions: a. DNA replication (3 minutes long) 1 © 2016 Carolina Biological Supply Company
b. Transcription and Translation (2 minutes long) c. Transcription and Translation 2 (5 minutes long) 2. What is a gene? Where does it start and where does it end? Are all genes within a cell transcribed and translated? - A gene is a fundamental component of heredity and a DNA sequence of nucleotides that codes for the production of an RNA or protein, depending on the gene. Codons for amino acids are used to begin genes, while stop codons are used to finish them. Not all of a cell's genes are translated and transcribed. Transcription is the initial stage of gene expression. It converts messenger RNA molecules into the genetic information contained in genes. It is subsequently transported to the cytoplasmic locations where proteins are synthesized. That's where translation happens. Reference: U.S. National Library of Medicine. (2021, March 22). What is a gene?: Medlineplus Genetics. MedlinePlus. Retrieved July 28, 2022, from https://medlineplus.gov/genetics/understanding/basics/gene/ 3. In our discussion forums we have been looking at how mutations can lead to various genetic diseases. One such genetic disease is sickle cell anemia that is the result of a mutation in the hemoglobin (Hb) beta subunit gene. In the table below is a partial sequence of the unmutated (Hb A) and mutated (Hb S) hemoglobin gene running 5’ to 3’. Please transcribe and translate each sequence making the mRNA and protein sequence of each. 2 © 2016 Carolina Biological Supply Company
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Molecule Sequence Hb A DNA 5’ GGC AGA CTT CTC CTC AGG AGT CAG GTG CAC 3’ Hb A mRNA 5’ CGC AGA CUU CUC AGG AGU CAG GUG CAC 3’ Hb A protein Gly-Arg-Leu-Leu-Leu-Arg-Ser-Gln-Val-His Hb S DNA 5’ GGC AGA CTT CTC CAC AGG AGT CAG GTG CAC 3’ Hb S mRNA 5’ GGC AGA CUU CUC CAC AGG AGU CAG GUG CAC 3’ Hb S protein Gly-Arg-Leu-Leu-His-Arg-Ser-Gln-Val-His 4. Considering the information above, what type of mutation (missense, nonsense, silent) is found within Hb S? - The mutilation that is found within Hb S is a missense. 5. Important! In activity 2 please take a picture once you complete step 9. This picture is not requested in the investigative manual, but it is below. Activity 1 Instructions: 1. Open the investigative manual. Locate all the needed materials supplied in the kit and those you will need to supply yourself. 2. Lay them out in your work area. 3. Read through the entire set of instructions found in the investigative manual for the activity to avoid making mistakes when you go to execute the experiment. 4. Once you have read through the instructions go back to step 1 and begin executing the experiment. 3 © 2016 Carolina Biological Supply Company
5. Please answer the questions below and/or append appropriate representations of data (photos, graphs, etc). REMEMBER don’t clean up until you have taken the appropriate photos of your experiment as described below. Photo 1 – Activity 1 Take a picture and insert the image(s) of your DNA strand from step 5. of the “DNA structure” section in activity 1 of the investigative manual: Activity 2 Instructions : 1. Open the investigative manual. Locate all the needed materials supplied in the kit and those you will need to supply yourself. 2. Lay them out in your work area. 3. Read through the entire set of instructions found in the investigative manual for the activity to avoid making mistakes when you go to execute the experiment. 4 © 2016 Carolina Biological Supply Company
4. Once you have read through the instructions go back to step 1 and begin executing the experiment. 5. Please answer the questions below and/or append appropriate representations of data (photos, graphs, etc). REMEMBER don’t clean up until you have taken the appropriate photos of your experiment as described below. Photo 1 – Activity 2 Take a picture and insert the image(s) of your partially replicated DNA strand from step 9. of the “DNA replication” section in activity 2 of the investigative manual: 1. What does the “pink” portion of this replication fork represent? Why is it needed? This replication fort's "pink" portion stands for the RNA primer. According to Study, this is where the DNA polymerase starts. Photo 2 – Activity 2 Take a picture and insert the image(s) of your fully replicated DNA strand from step 13. of the “DNA replication” section in activity 2 of the investigative manual: 5 © 2016 Carolina Biological Supply Company
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1. During replication, is the original DNA sequence maintained in the new strands? Yes, the sequence of the original DNA has been preserved in both of the new strands. The sequence of the original DNA is preserved in the copy that is made, which is called replication. This is due to the fact that the DNA polymerase enzyme, which is responsible for duplicating the DNA molecule, uses the first strand of DNA as a template in order to make the new strands of DNA. As a consequence of this, the new strands will be an exact replica of the first strand. 1. Is the replication of the leading strand continuous? How about the lagging strand? The replication of the leading strand starts with two parents DNA strands that are separated. Activity 3 Instructions: 1. Open the investigative manual. Locate all the needed materials supplied in the kit and those you will need to supply yourself. 2. Lay them out in your work area. 6 © 2016 Carolina Biological Supply Company
3. Read through the entire set of instructions found in the investigative manual for the activity to avoid making mistakes when you go to execute the experiment. 4. Once you have read through the instructions go back to step 1 and begin executing the experiment. 5. Please answer the questions below and/or append appropriate representations of data (photos, graphs, etc). REMEMBER don’t clean up until you have taken the appropriate photos of your experiment as described below. Photo 1 – Activity 3 Take a picture and insert the image(s) of your RNA strand from step 5. of the “Transcription” section in activity 1 of the investigative manual: Data Table 1 Molecule Sequence DNA Sequence 3’-TAC CAC GTA GAC TGA-5’ DNA Complement 5’- ATG GTG CAT CTG ACT-3’ RNA Complement 5’ AUG GUG CAU CUG ACU-3 7 © 2016 Carolina Biological Supply Company
Activity 4 Instructions: 1. Open the investigative manual. Locate all the needed materials supplied in the kit and those you will need to supply yourself. 2. Lay them out in your work area. 3. Read through the entire set of instructions found in the investigative manual for the activity to avoid making mistakes when you go to execute the experiment. 4. Once you have read through the instructions go back to step 1 and begin executing the experiment. 5. Please answer the questions below and/or append appropriate representations of data (photos, graphs, etc). REMEMBER don’t clean up until you have taken the appropriate photos of your experiment as described below. Data Table 2 Molecule Sequence Amino Acid MET VAL HIS LEU THR Activity 5 Instructions: 1. Open the investigative manual. Locate all the needed materials supplied in the kit and those you will need to supply yourself. 2. Lay them out in your work area. 3. Read through the entire set of instructions found in the investigative manual for the activity to avoid making mistakes when you go to execute the experiment. 8 © 2016 Carolina Biological Supply Company
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4. Once you have read through the instructions go back to step 1 and begin executing the experiment. 5. Please answer the questions below and/or append appropriate representations of data (photos, graphs, etc). REMEMBER don’t clean up until you have taken the appropriate photos of your experiment as described below. Data Table 3 Molecule Sequence Mutated DNA Sequence 3’ TAC CAC GTA AAC TGA- 5’ RNA Complement 5’ AUG GUG CAU UUC ACU-3 Amino Acid Sequence MET VAL HIS PHE THR 1. Explain the differences between the original amino acid sequence in the exercise and the one coded for by the mutated DNA/RNA. The start and finishing codons are different between the original amino acid sequence in the workout and the one encoded as altered DNA/RNA. Due to the two being altered, this would constitute a point mutation. 2. Will the protein translated from the mutated DNA function with the mutation that was generated? Explain your reasoning. Due to the mutation in the DNA the protein generation is difference in the non mutated DNA the protein generated is : Val His Leu Thr but in the mutated DNA sequence the amino acid sequence is : Glu His Leu Thr. 9 © 2016 Carolina Biological Supply Company
References: National human Genome Research Institute. “Gene.” Genome.gov, 2019, www.genome.gov/genetics-glossary/Gene. “Bioinformatics: Finding Genes.” Genome.gov, www.genome.gov/25020001/online-educationkit-bioinformatics-finding-genes#:~:text=A %20gene%20begins%20with%20a . 10 © 2016 Carolina Biological Supply Company