Lab report for crispr transfection

Rubric for Lab Report

These things must be present in your report to receive full points. (However, this is not a checklist and does not guarantee full points)

Title: You need a title, and your name. Format this like any paper.

Abstract: This should be a tiny version of your whole paper, less than 200 words.

Introduction: Start broad and work your way to a narrow point of view. Make sure you cover CRISPR and how that works. Go into a bit of details, and explain how it’s relevant. Compare CRISPR briefly to other gene insertion methods, and explain why CRISPR could be the best option for us. Mention RFP and puromycin resistance, and cover how they work. Talk about HEK 293T cells. Mention AAVS1 safe harbor site and what that is. Finally, Have a summary paragraph in which you state the purpose of our experiment, your hypothesis AND how you came to this hypothesis, how this was tested (the title of your key methods), and directly state your findings.

Materials and Methods: In paragraph form give a description for each technique used. This includes: The cell line used and their maintenance, the plasmids used, every experiment we performed, and the reagents used. If we made changes in class be sure you have those! This should be in paragraph form, and highly shortened. For example: Instead of saying “The media was put into a waste beaker and 3 mls of pbs was added and the pbs was taken off and put into the waste beaker. Then 10uL of puromycin was added to the cells and then media was added to those and left to incubate at 37 deg C for 48 hours” Simply say “The cells were washed and incubated in 15mM puromycin for 48 hours”. (Not actual concentration used)

Results: Express your results without giving raw data. This can be a bit tricky as data is a requirement. However, it should be expressed in words and references to the figures. For example: Instead of saying “Our groups BCA results had an optical density of 1.86” say this “the concentration of our extracted protein was 1865 ug/mL as determined by the BCA assay (Figure 3).” This requires you to understand how the experiments are related. However do not interpret your results.

You need 5 Figures with appropriate labels, a summary statement and a description all within the caption. These must be referenced in your text. Place your figures at the end of the lab report. They do not need to be in two columns, but your written results do.

The Figures Needed:

1. Both plasmids. You’ll need both plasmids correctly identified as guide or repair. In your caption identify key pieces of the plasmids.

2. Phalloidin and DAPI stain images. Label each stain present, as well as RFP. Also include a negative control.

3. The PCR agarose gel. You need to label key sizes of the ladder, each well, and label your band. Include the image of the PCR fragment we amplified

4. BCA standard curve as well as your concentration of protein

5. Western Blot of RFP in our CRISPR vs non CRISPR cells

Discussion: State if your hypothesis was supported or not, then explain why you believe this using your results. Go into details, and reference each figure while you discuss what they mean to our final conclusion. Next describe two ways in detail how we can further use our new skill. This should be at least a paragraph demonstrating you understand the concepts of what we did over the semester. Finally, have a conclusive ending statement.

References: Cite at least 5 Primary sources, just in your introduction. However, cite any background information you use. Use the Journal of Cell Biology format.

Formatting: The paper must be in two columns (excluding your abstract and references), single spaced, 12 point Times New Roman. Additionally use the paper on D2L as an example of how your paper should be formatted. Turn this in as a word document. Maximum of 6 pages (not including figures).

***Plagiarism: If your paper has plagiarism in any section, I will give you a zero. Depending on the extent of the plagiarism, I may turn you in to Dr. Jessen. Please be aware that the plagiarism detector on D2L is extremely sensitive and shows me the exact spot and sources of the plagiarism. Changing only a few words is still plagiarism, even if you cite the source. You must write in your own words.

************Rubric is subject to change as we discuss it. I will notify you of any changes in advance.