Purpose:

The purpose of this lab was to begin preparing for the CURE Symposium which will take place next Friday. We prepared our poster for printing and created abstracts that will be printed into the pamphlet for the event. Also during this lab we organized our soil metadata into a spreadsheet.

Material:

• Computer with internet connection.
• Previous notebooks.
• Soil Bag.
• Post-PCR DNA Sample.
• Poster.

Procedure:

1. Vote on logo for t-shirt and /or stickers.
2. Collect sample ID.
3. Enter metadata into spreadsheet and if PCR was positive, more it to the appropriate spot and place soil bag in container.
4. Go to the computer lab and review poster, and make necessary changes.
5. Discuss abstract and combine ideas to make final abstract.

Data:

Soil eDNA Metadata Spreadsheet:

Group Members: A.J. Alvarez, Megan Cordova, and Jessica Mann.

Section: 25

Group: 1

Soil ID: AA25_1Sp19

GPS Location: (-97.1136,31.5445)

Tree Species: unidentifiable

pH: 6.9

Soil Texture: Silty Loam

Extraction Method: Silica Bead

DNA Concentration ng/microleter: 479.5

Estimated Volume: 50

PCR: –

Soil Label on Bag: AA25SP19

Abstract:

Not much research has been done on the topic of soil ciliates. Therefore, a thorough research experiment on the study of soil ciliates’ DNA is necessary. This experiment was conducted to determine the diversity of ciliates from soil samples collected from the rhizosphere of different trees on the Baylor campus. We collected the metadata of the samples, such as tree and soil type, location, soil pH and percent water content. The silica bead eDNA extraction protocol and the Chelex DNA extraction from Ciliate culture were run on separate samples. We then ran gel electrophoresis, quantified the extracted DNA using Nanodrop and attempted to amplify the 18sV4 region using PCR. Gel electrophoresis yielded DNA bands for both extraction methods, however the gel run from the PCR reaction failed to produce results for either sample. Nanodrop showed that there were higher DNA concentrations for the silica bead extraction sample compared to the Chelex extraction samples. Due to the negative results for PCR amplification, DNA sequencing will not be performed on these samples.

Poster:

Conclusion:

In conclusion today’s lab had mostly to do with finalization. Since the semester is coming to an end, we are trying to prepare us for our final presentation next week. We finished up our posters as well as created our abstracts and I am feeling more comfortable when talking about the contents of my poster and the research we did this semester.

Future Steps:

In future labs I would like to practice presenting our poster and next Friday we will be presenting the research we did throughout the semester. I would also like to continue to run the metabarcoding so we can continue to look at the amazing diversity of Eukaryotas in the soil sample.