Question

hello

I need help with this i have to write a report about this

The importance of cell-cell contacts for cell function Professor Ellen Grandesco, principal investigator of the research group The Cell Contact Unit at the faculty of Medicine, LU, is in the process of writing a research proposal for a new project. The general aim of the proposal is to study the role of cell-cell contacts for the function of HEK293 cells, a human kidney cell line. Ellen needs preliminary data to show that she has an established cell system to base her functional cell studies on. To manipulate the cell-cell contacts of the HEK293 cells, Ellen decides to target E-Cadherin, a protein important for functional adherence junctions between cells. To modify the expression of E-Cadherin in the HEK293 cells Ellen purchases DNA vectors aimed at increasing or decreasing E-Cadherin expression. Hence, the following is available in the lab: HEK293 cells, two different transfection protocols and DNA vectors for manipulation of E-Cadherin expression.

1) Test if the supplied vectors affect the expression of E-Cadherin 2) Investigate which transfection method (of the two) works best with regard to transfection efficiency. Ellen points out that the transfection efficiency is important and she wants you to find out the following: a. How many of the cells are transfected with the respective transfection method expressed in % b. Is there a statistically significant difference between the transfection efficiencies of the two methods? 3) Investigate if manipulation of E-Cadherin expression affects the growth or shape of HEK293 cells. 4) Suggest and describe alternative methods to monitor the expression of E-Cadherin in this experiment (since only microscopy is available at this point in the laboratory). Include these suggestions in the discussion of your scientific report

1) Fluorescence microscope for cell culture plates 2) A 24-well plate with HEK293 cells, 150 000 cells/well 3) Two transfection protocols and reagents a. Lipofectamine 2000 (protocol below) b. PEI (protocol below) 4) DMEM (Dulbecco modified Eagle's minimal essential medium), Life Science; Catalogue number: SH30242.01 5) Penicillin/Streptomycin, Invitrogen; Catalogue number: 10378-016. This solution contains 10000 units of penicillin, 10000 µg of streptomycin, and 29.2 mg⁄ml of Lglutamine in a 10 mM citrate buffer (for pH stability). BIMA46 HT2019 Laborationshandledning 6 6) PBS (Phosphate Buffered Saline), Sigma-Aldrich; Catalogue number: D8537-500 ml 7) Opti-MEM I-Reduced serum medium, GIBCO; Catalogue number: 11058021 8) Trypsin-EDTA (0.25%), Thermo Fisher Scientific; Catalogue number: 25200056 9) FBS (Fetal Bovine serum), GIBCO. 10) DNA Vectors (0.5 µg DNA/µl) a. Scrambled GFP-ShRNA vector b. GFP-E-Cadherin vector c. GFP-E-Cadherin shRNA vector 11) Hoechst 33342, trihydrochloride, trihydrate (10 mg/mL solution in water)

The article should be less than 3000 words in total. There is no lower limit on the word count but all parts below have to be included and described properly. The required file format is MS Word. Content 1) Title page, including name and total word count 2) Abstract – one to two sentences about each of the following (max 250 words): a. Background b. Method c. Results d. Main conclusion (Write the abstract after you have finished writing your report) 3) Background and aim - describe the theoretical background and the scientific aim of the experiment (2-3 paragraphs). 4) Experimental method – write this section as you would find it described in a scientific article. This section usually has subheadings such as: Cell culture, Transfection, Microscopy, Statistical analysis. This section should ideally have a level of detail that enables a reader (an experienced scientist) to repeat the experiment. It should be written as running text, and thus not in bullet form (like protocols). 5) Results – describe the results and include references to the figures that are to be included in this section. The figures should be properly numbered and have a figure legend explaining what the figure represents. The figures may be attached to the end of the document or (preferably) inserted in the running text of the report. 6) Discussion of the results and the experiments’ strengths and weaknesses. The discussion should also describe and suggest alternative methods to investigate the expression of E-Cadherin in this experiment (since only microscopy is available at this point in the laboratory). 7) References – here you list references you have used in the introduction, material, result and discussion section of your report numbered in the order of appearance. The use of references in the report is required. Also, EndNote should be used to manage references.

resultat

lipo PEI

16% 12%

22% 13%

15%    12%

16% 10%

Answer 1

Proposal of a research work in the present era is a challenging task due to the constantly evolving trends in a quantitative research design and the need to incorporate medical advances into the methodology. The proposal is detailed plan or 'blueprint' for the intended study, and once it is completed, the research project should flow smoothly. Even today many of the proposals at post-graduate evaluation committee and application proposals for funding are substandard. A search was conducted with keywords such as research proposal, writing proposal and qualitative using search engine namely pubmed and google scholar, and an attempt has been made to provide broad guide lines for writing a scientifically appropriate research proposal.