Week 5: Lab and Lit

Welcome back to my blog!

This week I continued to read. I have continued reading silent spring by Rachel Carlson [1] and I read more about regulations regarding GMOs [2, 3].

In the lab:

On Tuesday, I made a DNA extraction for the petals of the transformed CRISPR arabidopsis then performed a PCR and let the samples incubate overnight.

The next day, I ran the PCR on a gel to make sure that the DNA extraction for the Arabidopsis flowers worked. It did, so I did a PCR purification and then tested those with a nanodrop to see the DNA concentration of the product. On Thursday, my mentor sent in those samples for sequencing, so we can confirm that the mutations are present.

On Friday after the lab meeting and seminar, My mentor and I used the overnight culture we set up on Wednesday to infiltrate some N. Benth (related to tobacco) leaves. To infiltrate the leaves we used syringes (without needles) and pressure to push the agrobacterium into the underside of the leaves. The reason why we used the underside of the leaf is that the underside has stomata (i.e. guard cells), this part of the plant is where gas exchange occurs. The stomata are basically like little mouths that can let the Agrobacterium into the plant. Next week will image them under the confocal microscope to look for plasmid localization within the plant cells. (The plasmid has a Green Fluorescent Protein (GFP) marker, so we will be looking for a green fluorescence signal during the screening.)

After infiltrating the leaves we went to the farm. There I worked on staking our first-generation transformed plants so that we can easily separate them when it is time to collect their seeds. Then I collected the last of the seeds from the floral dip Arabidopsis. Lastly, I helped my mentor with her maize organization.

Sources:

1. Silent Spring-Rachel Carson-1962: Free Download, borrow, and streaming. Internet Archive. (n.d.). Retrieved April 6, 2022, from https://archive.org/details/fp_Silent_Spring-Rachel_Carson-1962/page/n11/mode/1up
2. Agricultural Biotechnology Education and Outreach Initiative | FDA. (2022, February 17). US Food and Drug Administration. Retrieved April 25, 2022, from https://www.fda.gov/food/agricultural-biotechnology/agricultural-biotechnology-education-and-outreach-initiative
3. BE Disclosure | Agricultural Marketing Service. (n.d.). Agricultural Marketing Service. Retrieved April 25, 2022, from https://www.ams.usda.gov/rules-regulations/be