Objective

The objective of today’s lab was to work on our abstracts and finalize edits on our posters. We also needed to record all of our soil metadata and information on the excel sheet and label our soil sample with the correct identifier.

Purpose

The purpose of recording all our soil metadata was to have all of the class’s information in one spot so that a decision on which samples to send off for sequencing can be made. We then labeled our soil samples with a new identifier so they could be more easily found in storage to be sent off and so that it would match the identifier on the spreadsheet. We needed to work on our abstracts so there would be a preview of our presentation in the CURES pamphlet, and we needed to finalize our posters so they can be printed to be presented next week.

Procedure

1. Fill out Microsoft Excel sheet with soil information
2. Vote on options of logos
3. Retrieve soil sample and label it with “AHM22_5Sp19” and place it in the microfuge tube rack
4. Complete abstract with the lab group to turn in
5. Edit poster and upload to BOX to be printed
6. Upload abstract to box

Soil Metadata

Abstract/Title

Analysis of Soil and eDNA from the Rhizosphere of a Quercus virginia

The biodiversity of terrestrial ecosystems is invaluable to the ability for organisms to live on land. Soil microbiomes, located in these terrestrial ecosystems, are unexplored and the influence of eukaryotes on tree rhizospheres is unknown. This study analyzes soil collected on Baylor University’s campus from the rhizosphere of a Quercus virginiana to attempt to further classify ciliates and understand the diversity of one of the most controversial monophyletic groups, SAR. From the collected soil, metadata was collected for each sample including soil texture, percentage of water content, pH level, the circumference of tree collected from, and type of tree. The samples were then isolated and eDNA was extracted using silica beads. The eDNA was then cleaned and primed with an 18S V4 primer to isolate the V4 region of DNA in eukaryotes. PCR amplification was also done on the sample, and both were run through gel electrophoresis to attempt to view bands of DNA. Results from the PCR sequencing were analyzed using nanodrop technology and a gel imager. Both MHN and KRM samples yielded an A260/280 concentration of close to 1.8, indicating that DNA was present in the sample. Though it was shown that DNA was present, gel electrophoresis images showed no quantifiable results. Qiime2 was used to analyze positive samples and a decision on whether or not to send samples to a secondary lab for further sequencing was made.

Finalized Poster

Cleanup/Storage

The soil samples were placed back in the lab freezer to be stored. The area we were working at was cleaned up of all trash.

Future Goals

My future goal is to work on presentations and do well at the symposium. Our sample will not get sent off because we had negative results, so I don’t really have any goals to do with that. I think we have done as much as we can with our sample. Other than that, I have nothing because we’re finally done with this million-year-long semester.