DUE IN A DAY KINDLY HELP ME OUT
Q1. Suppose that you are a PhD student who works on stem cell differentiation. Your PI asked you to produce a large amount of BMP-2 protein to make osteogenic differentiation procedure faster. In order to produce large amount of protein, you were inserted 2 different candidate gene regions inserts with different lenghts into ampicilline resistant pGEM-bl vector to produce large amounts of BMP-2 protein by using E. coli Rosetta strain. After you inserted the plasmid and seed the bacteria in liquid agar, the government declared a 2-day quarantine due to Covid-19. Thereby you have an overpopulated bacteria solution and your PI wanted you to propose a long proposed experimental procedure to check whether they could be able to insert plasmids into bacteria to produce a BMP-2 protein and which insert candidate produce more BMP-2 protein via several steps: 1. Select the bacteria which has antibiotic resistant plasmid via seeding your bacteria solution by using proper procedure and materials (like proper mediums, agars etc.) and suppress the others. 2. Please note that, here, you have overpopulated bacteria solution. In order to perform further analysis, your colonies has to be countable and you have to be able to observe the morphology of these colonies and have to be able to select easily. (Please use hypertonic or hypotonic or isotonic solution if dilution is required and please explain why you are using your selected solution). 3. After selecting your bacteria which has antibiotic resistant plasmid, please propose 2 different methods to check whether the colonies are carrying your insert in the proper direction and use one of them by explaining the reasons why you select it. In here you have to check your bacteria's genome only. 4. Please compare these 2 different inserts that you obtained finally in terms of production rate of BMP-2 by designing new experiment. You're expecting to prepare a homework which includes an introduction, materials&methods, experimental procedure and proper expected results parts. The grading criteria for each part is following: a. Writing the procedure in detail. (35p) b. Proposing proper expected results for your experimental procedure.(20p) c. Writing an introduction to each technique that you plan to use in your procedure(35p) d. Giving at least 7 references in the AMA citation style. (10p) Q1. Suppose that you are a PhD student who works on stem cell differentiation. Your PI asked you to produce a large amount of BMP-2 protein to make osteogenic differentiation procedure faster. In order to produce large amount of protein, you were inserted 2 different candidate gene regions inserts with different lenghts into ampicilline resistant pGEM-bl vector to produce large amounts of BMP-2 protein by using E. coli Rosetta strain. After you inserted the plasmid and seed the bacteria in liquid agar, the government declared a 2-day quarantine due to Covid-19. Thereby you have an overpopulated bacteria solution and your PI wanted you to propose a long proposed experimental procedure to check whether they could be able to insert plasmids into bacteria to produce a BMP-2 protein and which insert candidate produce more BMP-2 protein via several steps: 1. Select the bacteria which has antibiotic resistant plasmid via seeding your bacteria solution by using proper procedure and materials (like proper mediums, agars etc.) and suppress the others. 2. Please note that, here, you have overpopulated bacteria solution. In order to perform further analysis, your colonies has to be countable and you have to be able to observe the morphology of these colonies and have to be able to select easily. (Please use hypertonic or hypotonic or isotonic solution if dilution is required and please explain why you are using your selected solution). 3. After selecting your bacteria which has antibiotic resistant plasmid, please propose 2 different methods to check whether the colonies are carrying your insert in the proper direction and use one of them by explaining the reasons why you select it. In here you have to check your bacteria's genome only. 4. Please compare these 2 different inserts that you obtained finally in terms of production rate of BMP-2 by designing new experiment. You're expecting to prepare a homework which includes an introduction, materials&methods, experimental procedure and proper expected results parts. The grading criteria for each part is following: a. Writing the procedure in detail. (35p) b. Proposing proper expected results for your experimental procedure.(20p) c. Writing an introduction to each technique that you plan to use in your procedure(35p) d. Giving at least 7 references in the AMA citation style. (10p)
