Lab 14

04/25/2019

Andreea Loghin

Objective:

The objective if this lab was to edit the poster and write the abstract for the Cures Symposium as well as contribute to the comprehensive spreadsheet with data about our DNA sample and tree and soil metadata.

Purpose:

The purpose of this lab was to give us a better understanding of what it means to participate in a scientific poster presentation and what the standards are for such posters as well as determine which DNA sample is viable to be sent for future analysis.

Procedures:

1. Lable vile with sample identifier (GLR22_7Sp19)
2. return tube to cooler
3. add data about latitude and longitude, tree diameter, pH, soil texture, methods of extraction, PCR results, DNA concentration, volume to the spreadsheet
4. work on poster
5. write abstract

Results:

SOIL eDNA metadata:

Group members: Sara Rothrock, Rebekah Gerry, Andreea Loghin

Section: 22

Group: 7

SOIL ID: GLR22_7Sp19

GPS location: -97.1195413, 31.5495435

Tree Species: Quercus virginiana

BHD (cm): 55.0064

pH: 6.5

Soil Texture: Loamy sand

Extraction Method: Silica Bead

Abstract: 

There are many standard methods for extracting Ciliate eDNA but not all are effective and cost efficient.

Ciliates are protozoa in the Kingdom Protista. They play many different roles within soil and aquatic ecosystems. There has been very little research done on ciliate behavior and interactions within the soil. Our study targeted ciliates within the rhizosphere. The rhizosphere plays a key role in the cycling of nutrients as well as plant growth. Ciliates shape the diversity of microorganisms within the rhizosphere, thus their presence is vital to a healthy ecosystem.

The purpose of this study was to investigate new methods for eDNA extraction of ciliates.

Creating this new method was achieved by researching different ways to extract eDNA. These different methods were then combined to maximize the amount of eDNA collected. Metadata was collected regarding the soil sample and the environment it was collected from. Gels were run to determine the amount of DNA collected. The samples that contained an abundance of DNA underwent PCR in order to amplify the strand so they could more easily be sequenced.

The samples were loamy sand, with a pH of 6.5. The concentration of DNA in the sample was 1125 ng/ul and the A260/A280 value was 1.33. The DNA sequenced not only belonged to ciliate species, but fungal and bacterial species as well.

These results gave reason to believe the use of charcoal and Silica beads was effective in the extraction of DNA.

Poster: click here

Future Goals:

In the future, we hope to get more data about the DNA samples that will be sent off for future analysis as well as present our poster during the CURES Symposium. We hope to contribute to the scientific community with valuable information regarding DNA extraction methods since we developed novel ways to do that.

Storage:

DNA samples were stored in the cooler after they were labeled. Everything was taken off the lab tables.