Write (12-15 pages) of GFP Report: Should include the following directions down below. Make sure you answer everything in your own words.You will have to work site every source you used.
▪Abstract 250-300 words
▪Background
– introduction to project and overall project goals
-Background information necessary to understand the project
-pGLO plasmid construct drawing and description
–Genes present in plasmid and purpose of each
–GFP gene original source and placement in plasmid
-ara expression system•amp resistance gene (bla) & purpose
–amp resistance gene (bla) & purpose
▪ procedure written as synopsis (no more than 2 pages)
▪Results
-Data table from transformation results
-Calculation of transformation frequency
-General data/observations on transformation
-General comments/observations on column chromatography
–Table with list of samples & volumes taken during extraction and purification process
-BSA standard curve data and graph
-Protein concentration calculations pre and post GFP samples
–Photos of gels clearly labeled with molecular weight standard information and labeled with what is in each lane
-Pre-stained gel (UV light) showing native GFP bands
-Stained gels showing GFP bands clearly labeled
–Estimation of Molecular weights of native GFP and presumptive denatured GFP bands
▪Discussion:
-Introduction to project goals
-Discussion of steps/observations in transformation process
–Discussion of reason for each transformation plate meaning of data on each one (one paragraph for each plate)
–Presentation of transformation frequency calculation and meaning of results
-Discussion of growth of cells for extraction/purification
-Discussion of extraction process
–Discussion of column purification process in relation to each of the buffers used
–Discussion of Bradford Assay for protein concentration, including standard curve preparation, preparation/dilution of samples, why dilutions were chosen, which dilution data were selected for calculations and why
-Comparison discussion of pre and post purification protein assays
–Discussion of protein assay data in relation to pre and post gel samples
–Discussion of pre and post assays in relation to purity of the samples at each stage
–Discussion of molecular weight of GFP based on native protein gel results; which band is the GFP and why?
–Discussion of molecular weight of GFP based on denatured protein gel results; which band is the GFP and why?
▪Conclusion
– Was the project successful overall? Did it meet the stated goals?
–Which elements of the data showed the project’s success (or lack of success)?
-What additional steps would need to be taken to complete this project?
*References included and crosschecked against the narrative
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